2-oxindole compounds

ABSTRACT

Oxindole compounds useful for the treatment of CCR(9) mediated conditions or diseases are provided.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/932,114 filed Jul. 17, 2020, which is a continuation of U.S. patentapplication Ser. No. 16/524,618 filed Jul. 29, 2019, now. U.S. Pat. No.10,759,789, which is a divisional of U.S. patent application Ser. No.15/914,900 filed Mar. 7, 2018, now U.S. Pat. No. 10,421,748, which is acontinuation of U.S. patent application Ser. No. 15/408,896 filed Jan.18, 2017, now abandoned, which is an application claiming benefit under35 U.S.C. § 119(e) of U.S. Provisional Application No. 62/280,969 filedJan. 20, 2016, each of which is herein incorporated by reference in itsentirety for all purposes.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH AND DEVELOPMENT

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REFERENCE TO A “SEQUENCE LISTING,” A TABLE, OR A COMPUTER PROGRAMLISTING APPENDIX SUBMITTED ON A COMPACT DISK

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BACKGROUND OF THE INVENTION

The present disclosure provides compounds and pharmaceuticalcompositions containing one or more of those compounds or theirpharmaceutically acceptable salts, that are effective in inhibiting thebinding or function of chemokines to the CCR9 chemokine receptor. Asantagonists or modulators of the CCR9 chemokine receptor, the compoundsand compositions can have utility in treating various immune disorderconditions and diseases.

Chemokines, also known as chemotactic cytokines, are a group of smallmolecular-weight proteins that are released by a wide variety of cellsand have a variety of biological activities. Chemokines attract varioustypes of cells of the immune system, such as macrophages, T cells,eosinophils, basophils and neutrophils, and cause them to migrate fromthe blood to various lymphoid and none-lymphoid tissues. They mediateinfiltration of inflammatory cells to sites of inflammation, and areresponsible for the initiation and perpetuation of many inflammationdiseases (reviewed in Schall, Cytokine, 3:165-183 (1991), Schall et al.,Curr. Opin. Immunol., 6:865-873 (1994)).

In addition to stimulating chemotaxis, chemokines can induce otherchanges in responsive cells, including changes in cell shape, granuleexocytosis, integrin up-regulation, formation of bioactive lipids (e.g.,leukotrienes), respiratory burst associated with leukocyte activation,cell proliferation, resistance to induction of apoptosis andangiogenesis. Thus, chemokines are early triggers of the inflammatoryresponse, causing inflammatory mediator release, chemotaxis andextravasation to sites of infection or inflammation. They are alsostimulators of a multitude of cellular processes that bear importantphysiological functions as well as pathological consequences.

Chemokines exert their effects by activating chemokine receptorsexpressed by responsive cells. Chemokine receptors are a class ofG-protein coupled receptors, also known as seven-transmembranereceptors, found on the surface of a wide variety of cell types such asleukocytes, endothelial cells, smooth muscle cells and tumor cells.

Chemokines and chemokine receptors are expressed by intrinsic renalcells and infiltrating cells during renal inflammation (Segerer et al.,J. Am. Soc. Nephrol., 11:152-76 (2000); Morii et al., J. DiabetesComplications, 17:11-5 (2003); Lloyd et al. J. Exp. Med., 185:1371-80(1997); Gonzalez-Cuadrado et al. Clin. Exp. Immunol., 106:518-22 (1996);Eddy & Giachelli, Kidney Int., 47:1546-57 (1995); Diamond et al., Am. J.Physiol., 266: F926-33 (1994)).

T lymphocyte (T cell) infiltration into the small intestine and colonhas been linked to the pathogenesis of Coeliac diseases, food allergies,rheumatoid arthritis, human inflammatory bowel diseases (IBD) whichinclude Crohn's disease and ulcerative colitis. Blocking trafficking ofrelevant T cell populations to the intestine can lead to an effectiveapproach to treat human IBD. More recently, chemokine receptor-9(CCR(9)) has been noted to be expressed on gut-homing T cells inperipheral blood, elevated in patients with small bowel inflammationsuch as Crohn's disease and coeliac disease. The only CCR(9) ligandidentified to date, TECK (thymus-expressed chemokine) is expressed inboth the small and large intestines and the ligand receptor pair is nowthought to play a pivotal role in the development of IBD. In particular,this pair mediates the migration of disease causing inflammatory cellsto the intestine. See for example, Zaballos et al., J. Immunol.,162(10):5671-5675 (1999); Kunkel et al., J. Exp. Med., 192(5):761-768(2000); Papadakis et al., J. Immunol., 165(9):5069-5076 (2000);Papadakis et al., Gastroenterology, 121(2):246-254 (2001); Campbell etal., J. Exp. Med., 195(1):135-141 (2002); Wurbel et al., Blood,98(9):2626-2632 (2001); and Uehara et al., J. Immunol, 168(6):2811-2819(2002); Rivera-Nieves et al., Gastroenterology, 2006 November;131(5):1518-29; and Kontoyiannis et al., J. Exp. Med., Vol. 196, Number12, Dec. 16, 2002. In addition CCR(9) bearing lymphocytes have been showto mediate the pathology of filariasis (lymphatic filarial disease) andinhibition of CCR(9) has been correlated with reduction of the pathologyassociated with such conditions. See for example Babu et al., Journal ofInfectious Diseases, 191: 1018-26, 2005.

The identification of compounds that modulate the function of CCR(9)represents an attractive new family of therapeutic agents for thetreatment of inflammatory and other conditions and diseases associatedwith CCR(9) activation, such as inflammatory bowel disease.

BRIEF SUMMARY OF THE INVENTION

The present invention is directed to compounds and pharmaceuticallyacceptable salts thereof, compositions, and methods useful in modulatingthe function of CCR(9). The compounds and salts thereof, compositions,and methods described herein can be useful in treating or preventingchemokine-mediated conditions or diseases, including certaininflammatory and immunoregulatory disorders and diseases.

The compounds of the present disclosure have been shown to modulateCCR(9), as shown in the examples.

In one aspect, the present compounds may be represented by formula (I):

or a pharmaceutically acceptable salt thereof, wherein

-   Ar is a 5- to 10-membered aromatic or heteroaromatic ring,    optionally substituted with from one to three R³;-   L¹ is selected from the group consisting of a bond, C₁₋₆ alkylene,    and C₁₋₆ heteroalkylene,-   L² is selected from the group consisting of a bond, C₁₋₆ alkylene,    and C₁₋₆ heteroalkylene,-   Y is CO₂H or a carboxylic acid bioisostere;-   each R¹ and each R^(2a) is independently selected from the group    consisting of halogen, cyano, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆    haloalkyl, C₁₋₆ haloalkoxy, C₃₋₆ cycloalkyl, and C₂₋₆ alkenyl,    wherein the alkyl, cycloalkyl and alkenyl portions are optionally    substituted with from one to three members selected from fluoro, OH,    CN, C₁₋₃ alkyl, C₁₋₃ haloalkyl, and C₁₋₃ alkoxy;-   R^(2b) is selected from the group consisting of H, halogen, cyano,    C₁₋₆ alkyl, C₁₋₆ alkoxy, C₃₋₆ cycloalkyl, and C₂₋₆ alkenyl, wherein    the alkyl, cycloalkyl and alkenyl portions are optionally    substituted with from one to three members selected from fluoro, CN,    C₁₋₃ alkyl, C₁₋₃ haloalkyl, and C₁₋₃ alkoxy;    -   or optionally one R^(2a) and R^(2b) when on adjacent vertices of        a phenyl ring, may be joined together to form a 5- or 6-membered        heterocycloalkyl ring having one or two ring vertices        independently selected from O, N and S, wherein said        heterocycloalkyl ring is optionally substituted with from one to        three members selected from fluoro and C₁₋₃ alkyl;-   each R³ is independently selected from the group consisting of    halogen, cyano, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkyl, C₁₋₆    haloalkoxy, C₃₋₆ cycloalkyl, and C₂₋₆ alkenyl;-   the subscript m is an integer of from 0 to 4; and-   the subscript n is an integer of from 0 to 3.

In another aspect, the present disclosure provides compositions usefulin modulating chemokine activity. In one embodiment, a compositionaccording to the present disclosure comprises a compound according tothe disclosure and a pharmaceutically acceptable carrier or excipient.

In yet another aspect, the present disclosure provides methods ofmodulating chemokine function in a cell, comprising contacting the cellwith a therapeutically effective amount of a compound or compositionaccording to the disclosure.

In still another aspect, the present disclosure provides methods formodulating chemokine function, comprising contacting a chemokinereceptor with a therapeutically effective amount of a compound orcomposition according to the disclosure.

In still another aspect, the present disclosure provides methods fortreating a chemokine-mediated condition or disease, comprisingadministering to a subject a safe and effective amount of a compound orcomposition according to the disclosure. The administering may be oral,parenteral, rectal, transdermal, sublingual, nasal or topical. In someaspects the compound may be administered in combination with ananti-inflammatory or analgesic agent.

In addition to the compounds provided herein, the present disclosurefurther provides pharmaceutical compositions containing one or more ofthese compounds, as well as methods for the use of these compounds intherapeutic methods, primarily to treat diseases associated withchemokine signaling activity. The CCR(9) mediated disease or conditionmay be inflammatory bowel diseases, an allergic disease, psoriasis,atopic dermatitis, asthma, fibrotic diseases, graft rejection, GvHD,Sjogren syndrome, immune mediated food allergies, autoimmune diseases,Celiac disease, rheumatoid arthritis, thymoma, thymic carcinoma,leukemia, solid tumor, or acute lymphocytic leukemia, melanoma, primarysclerosing cholangitis, hepatitis and inflammatory hepatic disease,post-operative ileus, Crohn's disease or ulcerative colitis.

BRIEF DESCRIPTION OF THE DRAWINGS

NOT APPLICABLE

DETAILED DESCRIPTION OF THE INVENTION General

The present disclosure is directed to compounds and salts thereof,compositions and methods useful in the modulation of chemokine receptorfunction, particularly CCR(9) function. Modulation of chemokine receptoractivity, as used herein in its various forms, is intended to encompassantagonism, agonism, partial antagonism, inverse agonism and/or partialagonism of the activity associated with a particular chemokine receptor,preferably the CCR(9) receptor. Accordingly, the compounds of thepresent disclosure are compounds which modulate at least one function orcharacteristic of mammalian CCR(9), for example, a human CCR(9) protein.The ability of a compound to modulate the function of CCR(9), can bedemonstrated in a binding assay (e.g., ligand binding or agonistbinding), a chemotaxis (migration assay), a signaling assay (e.g.,activation of a mammalian G protein, induction of rapid and transientincrease in the concentration of cytosolic free calcium), and/orcellular response assay (e.g., stimulation of chemotaxis, exocytosis orinflammatory mediator release by leukocytes).

Abbreviations and Definitions

The term “alkyl”, by itself or as part of another substituent, means,unless otherwise stated, a straight or branched chain hydrocarbonradical, having the number of carbon atoms designated (i.e. C₁₋₈ meansone to eight carbons). Examples of alkyl groups include methyl, ethyl,n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl,n-hexyl, n-heptyl, n-octyl, and the like. The term “alkenyl” refers toan unsaturated alkyl group having one or more double bonds. Similarly,the term “alkynyl” refers to an unsaturated alkyl group having one ormore triple bonds. Examples of such unsaturated alkyl groups includevinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl),2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl,3-butynyl, and the higher homologs and isomers. The term “cycloalkyl”refers to hydrocarbon rings having the indicated number of ring atoms(e.g., C₃₋₆cycloalkyl) and being fully saturated or having no more thanone double bond between ring vertices. “Cycloalkyl” is also meant torefer to bicyclic and polycyclic hydrocarbon rings such as, for example,bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, etc. The term“heterocycloalkane” or “heterocycloalkyl” refers to a cycloalkyl groupthat contain from one to five heteroatoms selected from N, O, and S,wherein the nitrogen and sulfur atoms are optionally oxidized, and thenitrogen atom(s) are optionally quaternized. The heterocycloalkane maybe a monocyclic, a bicyclic or a polycylic ring system. Non limitingexamples of heterocycloalkane groups include pyrrolidine, imidazolidine,pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin,dioxolane, phthalimide, piperidine, 1,4-dioxane, morpholine,thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide,piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone,tetrahydrofuran, tetrhydrothiophene, quinuclidine, and the like. Aheterocycloalkane group can be attached to the remainder of the moleculethrough a ring carbon or a heteroatom.

The term “alkylene” by itself or as part of another substituent means adivalent radical derived from an alkane, as exemplified by—CH₂CH₂CH₂CH₂—. Typically, an alkyl (or alkylene) group will have from 1to 24 carbon atoms, with those groups having 10 or fewer carbon atomsbeing preferred in the present disclosure. A “lower alkyl” or “loweralkylene” is a shorter chain alkyl or alkylene group, generally havingfour or fewer carbon atoms. Similarly, “alkenylene” and “alkynylene”refer to the unsaturated forms of “alkylene” having double or triplebonds, respectively. The term “heteroalkylene” refers to an alkylenegroup in which one or two carbon atoms are replaced by N, O, or S.

As used herein, a wavy line, “

”, that intersects a single, double or triple bond in any chemicalstructure depicted herein, represent the point attachment of the single,double, or triple bond to the remainder of the molecule.

The terms “alkoxy,” “alkylamino” and “alkylthio” (or thioalkoxy) areused in their conventional sense, and refer to those alkyl groupsattached to the remainder of the molecule via an oxygen atom, an aminogroup, or a sulfur atom, respectively. Additionally, for dialkylaminogroups, the alkyl portions can be the same or different and can also becombined to form a 3-7 membered ring with the nitrogen atom to whicheach is attached. Accordingly, a group represented as dialkylamino or—NR^(a)R^(b) is meant to include piperidinyl, pyrrolidinyl, morpholinyl,azetidinyl and the like.

The term “di-(C₁₋₄alkyl)amino-C₁₋₄ alkyl” refers to an amino groupbearing two C₁₋₄ alkyl groups that can be the same or different (e.g.,methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl andtert-butyl) and which is attached to the remainder of the moleculethrough a C₁₋₄ alkyl group (a one to four carbon alkylene linkinggroup). Examples of di-(C₁₋₄ alkyl)amino-C₁₋₄ alkyl groups includedimethylaminomethyl, 2-(ethyl(methyl)amino)ethyl,3-(dimethylamino)butyl, and the like.

The terms “halo” or “halogen,” by themselves or as part of anothersubstituent, mean, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom. Additionally, terms such as “haloalkyl” and“haloalkoxy,” are meant to include monohalo- and polyhalo-versions ofalkyl and alkoxy, respectively. For example, the term “C₁₋₄haloalkyl” ismean to include trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl,3-bromopropyl, and the like.

The term “aryl” or “aromatic ring” means, unless otherwise stated, apolyunsaturated, typically aromatic, hydrocarbon group which can be asingle ring or multiple rings (up to three rings) which are fusedtogether or linked covalently. Similarly, the terms “heteroaryl” and“heteroaromatic ring” refer to aryl groups (or rings) that contain fromone to five heteroatoms selected from N, O, and S, wherein the nitrogenand sulfur atoms are optionally oxidized, and the nitrogen atom(s) areoptionally quaternized. A heteroaryl group or heteroaromatic ring can beattached to the remainder of the molecule through a heteroatom.Non-limiting examples of aryl groups include phenyl, naphthyl andbiphenyl, while non-limiting examples of heteroaryl groups includepyridyl, pyridazinyl, pyrazinyl, pyrimindinyl, triazinyl, quinolinyl,quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, benzotriazinyl,purinyl, benzimidazolyl, benzopyrazolyl, benzotriazolyl, benzisoxazolyl,isobenzofuryl, isoindolyl, indolizinyl, benzotriazinyl, thienopyridinyl,thienopyrimidinyl, pyrazolopyrimidinyl, imidazopyridines,benzothiaxolyl, benzofuranyl, benzothienyl, indolyl, quinolyl,isoquinolyl, isothiazolyl, pyrazolyl, indazolyl, pteridinyl, imidazolyl,triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrrolyl,thiazolyl, furyl, thienyl and the like. Substituents for each of theabove noted aryl and heteroaryl ring systems are selected from the groupof acceptable substituents described below.

The term “arylalkyl” is meant to include those radicals in which an arylgroup is attached to an alkyl group (e.g., benzyl, phenethyl, and thelike). Similarly, the term “heteroaryl-alkyl” is meant to include thoseradicals in which a heteroaryl group is attached to an alkyl group(e.g., pyridylmethyl, thiazolylethyl, and the like).

As used herein, the term “heteroatom” is meant to include oxygen (O),nitrogen (N), sulfur (S) and silicon (Si).

The term “carboxylic acid bioisostere” refers to a group having polarand/or acidic character to act as a replacement for a carboxylic acid. Avariety of bioisosteres are known for carboxylic acids, including,hydroxamic acids, hydroxamic esters, phosphonic acids, phosphinic acids,sulfonic acids, sulfinic acids, sulfonamides, acyl sulfonamides,acylureas, sulfonylureas, cyclopentane-1,2-diones, substituted phenols,and heterocycle-based bioisosteres as provided below:

wherein p is 0, 1 or 2 and wherein each R group is independentlyselected from the group consisting of H, C₁-C₆ alkyl, C₁-C₆ haloalkyl,C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy, or C₁-C₄ alkyl-O—C₁-C₄ alkyl.

Other examples of bioisosteres of a carboxylic acid are a tetrazolyl ortetrazolonyl, wherein the tetrazolyl or tetrazolonyl is optionallysubstituted with C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ hydroxyalkyl, —C₁₋₆alkoxy or C₁₋₄ alkyl-O—C₁₋₄ alkyl. Still other examples of carboxylicacid bioisosteres are described in the Journal of Medicinal Chemistry,2016, 59, 3183-3203 which are hereby incorporated by reference.

The term “pharmaceutically acceptable salts” is meant to include saltsof the active compounds which are prepared with relatively nontoxicacids or bases, depending on the particular substituents found on thecompounds described herein. When compounds of the present disclosurecontain relatively acidic functionalities, base addition salts can beobtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of salts derived frompharmaceutically-acceptable inorganic bases include aluminum, ammonium,calcium, copper, ferric, ferrous, lithium, magnesium, manganic,manganous, potassium, sodium, zinc and the like. Salts derived frompharmaceutically-acceptable organic bases include salts of primary,secondary and tertiary amines, including substituted amines, cyclicamines, naturally-occurring amines and the like, such as arginine,betaine, caffeine, choline, N,N′-dibenzylethylenediamine, diethylamine,2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperidine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylamine,tripropylamine, tromethamine and the like. When compounds of the presentdisclosure contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, malonic, benzoic, succinic, suberic,fumaric, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric,tartaric, methanesulfonic, and the like. Also included are salts ofamino acids such as arginate and the like, and salts of organic acidslike glucuronic or galactunoric acids and the like (see, for example,Berge, S. M., et al, “Pharmaceutical Salts”, Journal of PharmaceuticalScience, 1977, 66, 1-19). Certain specific compounds of the presentdisclosure contain both basic and acidic functionalities that allow thecompounds to be converted into either base or acid addition salts.

The neutral forms of the compounds may be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents, but otherwise the salts are equivalent to the parentform of the compound for the purposes of the present disclosure.

In addition to salt forms, the present disclosure provides compoundswhich are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentdisclosure. Additionally, prodrugs can be converted to the compounds ofthe present disclosure by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the present disclosure when placed in a transdermal patchreservoir with a suitable enzyme or chemical reagent.

Certain compounds of the present disclosure can exist in unsolvatedforms as well as solvated forms, including hydrated forms. In general,the solvated forms are equivalent to unsolvated forms and are intendedto be encompassed within the scope of the present disclosure. Certaincompounds of the present disclosure may exist in multiple crystalline oramorphous forms. In general, all physical forms are equivalent for theuses contemplated by the present disclosure and are intended to bewithin the scope of the present disclosure.

Certain compounds of the present disclosure possess asymmetric carbonatoms (optical centers) or double bonds; the racemates, diastereomers,geometric isomers, regioisomers and individual isomers (e.g., separateenantiomers) are all intended to be encompassed within the scope of thepresent disclosure. When a stereochemical depiction is shown, it ismeant to refer the compound in which one of the isomers is present andsubstantially free of the other isomer. ‘Substantially free of’ anotherisomer indicates at least an 80/20 ratio of the two isomers, morepreferably 90/10, or 95/5 or more. In some embodiments, one of theisomers will be present in an amount of at least 99%.

The compounds of the present disclosure may also contain unnaturalproportions of atomic isotopes at one or more of the atoms thatconstitute such compounds. Unnatural proportions of an isotope may bedefined as ranging from the amount found in nature to an amountconsisting of 100% of the atom in question. For example, the compoundsmay incorporate radioactive isotopes, such as for example tritium (³H),iodine-125 (¹²⁵I) or carbon-14 (¹⁴C), or non-radioactive isotopes, suchas deuterium (²H) or carbon-13 (¹³C). Such isotopic variations canprovide additional utilities to those described elsewhere with thisapplication. For instance, isotopic variants of the compounds of thedisclosure may find additional utility, including but not limited to, asdiagnostic and/or imaging reagents, or as cytotoxic/radiotoxictherapeutic agents. Additionally, isotopic variants of the compounds ofthe disclosure can have altered pharmacokinetic and pharmacodynamiccharacteristics which can contribute to enhanced safety, tolerability orefficacy during treatment. All isotopic variations of the compounds ofthe present disclosure, whether radioactive or not, are intended to beencompassed within the scope of the present disclosure. Substitutionwith heavier isotopes such as deuterium, i.e. ²H, may afford certaintherapeutic advantages resulting from greater metabolic stability. Forexample, in vivo half-life may increase or dosage requirements may bereduced.

Compounds of the disclosure having formula I can exist in differentisomeric forms. As used herein, the terms cis or trans are used in theirconventional sense in the chemical arts, i.e., referring to the positionof the substituents to one another relative to a reference plane, e.g.,a double bond, or a ring system, such as a decalin-type ring system or ahydroquinolone ring system: in the cis isomer, the substituents are onthe same side of the reference plane, in the trans isomer thesubstituents are on opposite sides. Additionally, different conformersare contemplated by the present disclosure, as well as distinctrotamers. Conformers are conformational isomers that can differ byrotations about one or more 6 bonds. Rotamers are conformers that differby rotation about only a single a bond.

Compounds

The present disclosure provides compounds that modulate the activity ofCCR(9). Chemokine receptors are integral membrane proteins whichinteract with an extracellular ligand, such as a chemokine, and mediatea cellular response to the ligand, e.g., chemotaxis, increasedintracellular calcium ion concentration, etc. Therefore, modulation of achemokine receptor function, e.g., interference with a chemokinereceptor ligand interaction, will modulate a chemokine receptor mediatedresponse, and treat or prevent a chemokine receptor mediated conditionor disease. Modulation of a chemokine receptor function includes bothinducement and inhibition of the function. The type of modulationaccomplished will depend on the characteristics of the compound, i.e.,antagonist or full, partial or inverse agonist.

For example, compounds of this disclosure act as potent CCR(9)antagonists, and this antagonistic activity has been further confirmedin animal testing for inflammation, one of the hallmark disease statesfor CCR(9). Accordingly, the compounds provided herein are useful inpharmaceutical compositions, methods for the treatment ofCCR(9)-mediated diseases, and as controls in assays for theidentification of competitive CCR(9) antagonists.

Provided herein are compounds of formula (I):

or a pharmaceutically acceptable salt thereof, wherein

-   Ar is a 5- to 10-membered aromatic or heteroaromatic ring,    optionally substituted with from one to three R³;-   L¹ is selected from the group consisting of a bond, C₁₋₆ alkylene,    and C₁₋₆ heteroalkylene,-   L² is selected from the group consisting of a bond, C₁₋₆ alkylene,    and C₁₋₆ heteroalkylene,-   Y is CO₂H or a carboxylic acid bioisostere;-   each R¹ and each R^(2a) is independently selected from the group    consisting of halogen, cyano, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆    haloalkyl, C₁₋₆ haloalkoxy, C₃₋₆ cycloalkyl, and C₂₋₆ alkenyl,    wherein the alkyl, cycloalkyl and alkenyl portions are optionally    substituted with from one to three members selected from fluoro, OH,    CN, C₁₋₃ alkyl, C₁₋₃ haloalkyl, and C₁₋₃ alkoxy;-   R^(2b) is selected from the group consisting of H, halogen, cyano,    C₁₋₆ alkyl, C₁₋₆ alkoxy, C₃₋₆ cycloalkyl, and C₂₋₆ alkenyl, wherein    the alkyl, cycloalkyl and alkenyl portions are optionally    substituted with from one to three members selected from fluoro, CN,    C₁₋₃ alkyl, C₁₋₃ haloalkyl, and C₁₋₃ alkoxy;-   or optionally one R^(2a) and R^(2b) when on adjacent vertices of a    phenyl ring, may be joined together to form a 5- or 6-membered    heterocycloalkyl ring having one or two ring vertices independently    selected from O, N and S, wherein said heterocycloalkyl ring is    optionally substituted with from one to three members selected from    fluoro and C₁₋₃ alkyl;-   each R³ is independently selected from the group consisting of    halogen, cyano, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkyl, C₁₋₆    haloalkoxy, C₃₋₆ cycloalkyl, and C₂₋₆ alkenyl;-   the subscript m is an integer of from 0 to 4; and-   the subscript n is an integer of from 0 to 3.

In some embodiments, Y is selected from the group consisting of:

tetrazolyl and tetrazolonyl, wherein the tetrazolyl or tetrazolonyl isoptionally substituted with R, wherein p is 0, 1 or 2 and wherein each Rgroup is independently selected from the group consisting of H, C₁-C₆alkyl, C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy, or C₁-C₄alkyl-O—C₁-C₄ alkyl.

In some embodiments, Y is selected from the group consisting of:tetrazolyl and tetrazolonyl, wherein the tetrazolyl or tetrazolonyl isoptionally substituted with C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆hydroxyalkyl, —C₁₋₆ alkoxy or C₁₋₄alkyl-O—C₁₋₄alkyl.

In some embodiments, a compound is provided having the formula selectedfrom the group consisting of:

or a pharmaceutically acceptable salt thereof, wherein said compound issubstantially free of other isomers.

In some embodiments, a compound is provided having the formula:

or a pharmaceutically acceptable salt thereof, wherein L² is C₁₋₃alkylene and wherein said compound is substantially free of otherisomers.

In some embodiments, a compound is provided having the formula (I′):

or a pharmaceutically acceptable salt thereof, wherein

-   Ar is a 5- to 10-membered aromatic or heteroaromatic ring,    optionally substituted with from one to three R³;-   L¹ is selected from the group consisting of a bond, C₁₋₆ alkylene,    and C₁₋₆ heteroalkylene,-   L² is selected from the group consisting of a bond, C₁₋₆ alkylene,    and C₁₋₆ heteroalkylene,-   each R¹ and each R^(2a) is independently selected from the group    consisting of halogen, cyano, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆    haloalkoxy, C₃₋₆ cycloalkyl, and C₂₋₆ alkenyl, wherein the alkyl,    cycloalkyl and alkenyl portion are optionally substituted with from    one to three members selected from fluoro, CN, C₁₋₃ alkyl, C₁₋₃    haloalkyl, and C₁₋₃ alkoxy;-   R^(2b) is selected from the group consisting of H, halogen, cyano,    C₁₋₆ alkyl, C₁₋₆ alkoxy, C₃₋₆ cycloalkyl, and C₂₋₆ alkenyl, wherein    the alkyl, cycloalkyl and alkenyl portion are optionally substituted    with from one to three members selected from fluoro, CN, C₁₋₃ alkyl,    C₁₋₃ haloalkyl, and C₁₋₃ alkoxy;    -   or optionally one R^(2a) and R^(2b) when on adjacent vertices of        a phenyl ring, may be joined together to form a 5- or 6-membered        cycloheteroalkyl ring having one or two ring vertices        independently selected from O, N and S, wherein said        cycloheteroalkyl ring is optionally substituted with from one to        three members selected from fluoro and C₁₋₃ alkyl;-   each R³ is independently selected from the group consisting of    halogen, cyano, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkyl, C₁₋₆    haloalkoxy, C₃₋₆ cycloalkyl, and C₂₋₆ alkenyl;-   the subscript m is an integer of from 0 to 4; and-   the subscript n is an integer of from 0 to 3.

In one group of embodiments for each of formulae (I) and (I′), Ar isselected from benzene, pyridine and quinoline, each of which isoptionally substituted with from one to two R³.

In some selected embodiments of formula (I) and (I′), L¹ is selectedfrom the group consisting of a bond, —CH₂— and —CH(CH₃)—. In otherselected embodiments of formula (I), L² is selected from the groupconsisting of a

bond, —O—CH₂—, —CH(CH₃)—, —C(CH₃)₂—, —CH₂CH₂—, —CH₂—, and —CH₂CH₂CH₂—.

In some selected embodiments of formula (I) and (I′), n is 1 or 2. Inother selected embodiments of formula (I) and (I′), m is 1, 2 or 3.

In other embodiments, suitable compounds are provided having the formula(Ia):

or a pharmaceutically acceptable salt thereof.

In one group of selected embodiments, Ar is selected from benzene,pyridine and quinoline, each of which is optionally substituted withfrom one to two R³.

In another group of selected embodiments, L¹ is selected from the groupconsisting of a bond, —CH₂— and —CH(CH₃)—. In still another group ofselected embodiments L² is selected from the group consisting of a bond,—O—CH₂—, —CH(CH₃)—, —C(CH₃)₂—, —CH₂CH₂—, —CH₂—, and —CH₂CH₂CH₂—.

In other selected embodiments, suitable compounds are selected from:

or a pharmaceutically acceptable salt thereof, wherein said compound issubstantially free of other isomers.

Within formula (Ia1), (Ia2) and (Ia3), selected embodiments are thosewherein Ar is selected from the group consisting of benzene, pyridineand quinoline, each of which is optionally substituted with from one totwo R³. In still other embodiments, Ar is selected from the groupconsisting of 1,3-phenylene and 1,4-phenylene, each of which isoptionally substituted with from one to two R³. In some embodiments,referring to formulae (Ia1), (Ia2) and (Ia3), L¹ is selected from thegroup consisting of a bond, —CH₂— and —CH(CH₃)—. In other embodiments offormulae (Ia1), (Ia2) and (Ia3), L² is selected from the groupconsisting of —O—CH₂—, —CH(CH₃)—, —C(CH₃)₂—, —CH₂CH₂—, —CH₂— and—CH₂CH₂CH₂—. In still other embodiments of formulae (Ia1), (Ia2) and(Ia3), le is selected from the group consisting of halogen, cyano, C₁₋₃alkyl, C₁₋₃ alkoxy, C₁₋₃ haloalkyl, C₁₋₃ haloalkoxy, C₃₋₅ cycloalkyl,and C₂₋₃ alkenyl. In yet other embodiments of formulae (Ia1), (Ia2) and(Ia3), R¹ is selected from the group consisting of chloro, methyl,cyano, ethyl, cyclopropyl, trifluoromethyl and trifluoromethoxy.

In other selected embodiments, suitable compounds are selected from:

or a pharmaceutically acceptable salt thereof, wherein said compound issubstantially free of other isomers.

In other embodiments, suitable compounds are provided having the formula(Ib):

or a pharmaceutically acceptable salt thereof.

In some embodiments of formula (Ib), R^(2b) is hydrogen. In otherembodiments of formula (Ib), Ar is selected from the group consisting ofbenzene, pyridine and quinoline, each of which is optionally substitutedwith from one to two R³. In still other embodiments of formula (Ib), L¹is selected from the group consisting of a bond, —CH₂— and —CH(CH₃)—. Inyet other embodiments of formula (Ib), L² is selected from the groupconsisting of a bond, —O—CH₂—, —CH(CH₃)—, —C(CH₃)₂—, —CH₂CH₂—, —CH₂— and—CH₂CH₂CH₂—.

In other selected embodiments, suitable compounds are selected from:

or a pharmaceutically acceptable salt thereof, wherein said compound issubstantially free of other isomers.

Within formula (Ib1), (Ib2) and (Ib3), selected embodiments are thosewherein Ar is selected from the group consisting of benzene, pyridineand quinoline, each of which is optionally substituted with from one totwo R³. In some embodiments, referring to formulae (Ib1), (Ib2) and(Ib3), Ar is selected from the group consisting of 1,3-phenylene and1,4-phenylene, each of which is optionally substituted with from one totwo R³. In other embodiments of formulae (Ib1), (Ib2) and (Ib3), R³ isselected from the group consisting of CH₃, CH₂CH₃, CH₂CH₂CH₃, CH(CH₃)₂and CH₂OH. In still other embodiments of formulae (Ib1), (Ib2) and(Ib3), L¹ is selected from the group consisting of a bond, —CH₂— and—CH(CH₃)—. In yet other embodiments of formulae (Ib1), (Ib2) and (Ib3),L² is selected from the group consisting of —O—CH₂—, —CH(CH₃)—,—C(CH₃)₂—, —CH₂CH₂—, —CH₂— and —CH₂CH₂CH₂—. In other embodiments offormulae (Ib1), (Ib2) and (Ib3), le is selected from the groupconsisting of halogen, cyano, C₁₋₃ alkyl, C₁₋₃ alkoxy, C₁₋₃ haloalkoxy,C₃₋₅ cycloalkyl, and C₂₋₃ alkenyl, or R¹ is selected from the groupconsisting of chloro, methyl, cyano, ethyl, cyclopropyl, trifluoromethyland trifluoromethoxy.

In other selected embodiments, suitable compounds are selected from:

or a pharmaceutically acceptable salt thereof, wherein said compound issubstantially free of other isomers.

Preparation of Compounds

Compound provided herein can be prepared by the general scheme below.Beginning with a suitably substituted phenylacetic acid ester, reactionwith a substituted halonitrobenzene in the presence of base, followingby methyl iodide sets the framework for 2-oxoindole ring constructionhaving the quaternary center alpha to the carboxylic acid. Resolution ofthe isomers, followed by reduction of the nitro group and cyclizationproduces a substituted 2-oxoindole. Reaction at the indole nitrogen atomto attach either a substituted Ar group, or a linker (L¹), having anattached substituted Ar group will lead to the target compounds shown.One of skill in the art will appreciate that modifications can be madefollowing the general guidance of the scheme below to provide a varietyof compounds of formula (I).

Compositions that Modulate Chemokine Activity

In another aspect, the present disclosure provides compositions thatmodulate chemokine activity, specifically CCR(9) activity. Generally,the compositions for modulating chemokine receptor activity in humansand animals will comprise a pharmaceutically acceptable excipient ordiluent and a compound having any of the formulae I, I′, Ia, Ib, Ia1,Ia2, Ia3, Ia1′, Ia2′, Ia3′, Ib1, Ib2, Ib3, Ib1 Ib2′ and Ib3′.

The term “composition” as used herein is intended to encompass a productcomprising the specified ingredients in the specified amounts, as wellas any product which results, directly or indirectly, from combinationof the specified ingredients in the specified amounts. By“pharmaceutically acceptable” it is meant the carrier, diluent orexcipient must be compatible with the other ingredients of theformulation and not deleterious to the recipient thereof.

The pharmaceutical compositions for the administration of the compoundsof this disclosure may conveniently be presented in unit dosage form andmay be prepared by any of the methods well known in the art of pharmacy.All methods include the step of bringing the active ingredient intoassociation with the carrier which constitutes one or more accessoryingredients. In general, the pharmaceutical compositions are prepared byuniformly and intimately bringing the active ingredient into associationwith a liquid carrier or a finely divided solid carrier or both, andthen, if necessary, shaping the product into the desired formulation. Inthe pharmaceutical composition the active object compound is included inan amount sufficient to produce the desired effect upon the process orcondition of diseases.

The pharmaceutical compositions containing the active ingredient may bein a form suitable for oral use, for example, as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules,emulsions and self-emulsifications as described in U.S. Pat. No.6,451,339, hard or soft capsules, or syrups or elixirs. Compositionsintended for oral use may be prepared according to any method known tothe art for the manufacture of pharmaceutical compositions. Suchcompositions may contain one or more agents selected from sweeteningagents, flavoring agents, coloring agents and preserving agents in orderto provide pharmaceutically elegant and palatable preparations. Tabletscontain the active ingredient in admixture with other non-toxicpharmaceutically acceptable excipients which are suitable for themanufacture of tablets. These excipients may be, for example, inertdiluents such as cellulose, silicon dioxide, aluminum oxide, calciumcarbonate, sodium carbonate, glucose, mannitol, sorbitol, lactose,calcium phosphate or sodium phosphate; granulating and disintegratingagents, for example, corn starch, or alginic acid; binding agents, forexample PVP, cellulose, PEG, starch, gelatin or acacia, and lubricatingagents, for example magnesium stearate, stearic acid or talc. Thetablets may be uncoated or they may be coated enterically or otherwiseby known techniques to delay disintegration and absorption in thegastrointestinal tract and thereby provide a sustained action over alonger period. For example, a time delay material such as glycerylmonostearate or glyceryl distearate may be employed. They may also becoated by the techniques described in the U.S. Pat. Nos. 4,256,108;4,166,452; and U.S. Pat. No. 4,265,874 to form osmotic therapeutictablets for control release.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example peanut oil, liquid paraffin, or olive oil.Additionally, emulsions can be prepared with a non-water miscibleingredient such as oils and stabilized with surfactants such asmono-diglycerides, PEG esters and the like.

Aqueous suspensions contain the active materials in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose,sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxyethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethyleneoxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl, p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present.

The pharmaceutical compositions of the disclosure may also be in theform of oil in water emulsions. The oily phase may be a vegetable oil,for example olive oil or arachis oil, or a mineral oil, for exampleliquid paraffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring gums, for example gum acacia or gum tragacanth,naturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative, and flavoring and coloringagents. Oral solutions can be prepared in combination with, for example,cyclodextrin, PEG and surfactants.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleaginous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents which have been mentioned above.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally acceptable diluent orsolvent, for example as a solution in 1,3-butane diol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, axed oils are conventionally employed as a solvent orsuspending medium. For this purpose any bland fixed oil may be employedincluding synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid find use in the preparation of injectables.

The compounds of the present disclosure may also be administered in theform of suppositories for rectal administration of the drug. Thesecompositions can be prepared by mixing the drug with a suitablenon-irritating excipient which is solid at ordinary temperatures butliquid at the rectal temperature and will therefore melt in the rectumto release the drug. Such materials are cocoa butter and polyethyleneglycols. Additionally, the compounds can be administered via oculardelivery by means of solutions or ointments. Still further, transdermaldelivery of the subject compounds can be accomplished by means ofiontophoretic patches and the like.

For topical use, creams, ointments, jellies, solutions or suspensionscontaining the compounds of the present disclosure are employed. As usedherein, topical application is also meant to include the use of mouthwashes and gargles.

The pharmaceutical compositions and methods of the present disclosuremay further comprise other therapeutically active compounds as notedherein, such as those applied in the treatment of the above mentionedpathological conditions.

In one embodiment, the present disclosure provides a compositionconsisting of a pharmaceutically acceptable carrier and a compound ofthe disclosure.

Methods of Treatment

Depending on the disease to be treated and the subject's condition, thecompounds and compositions of the present disclosure may be administeredby oral, parenteral (e.g., intramuscular, intraperitoneal, intravenous,ICV, intracisternal injection or infusion, subcutaneous injection, orimplant), inhalation, nasal, vaginal, rectal, sublingual, or topicalroutes of administration and may be formulated, alone or together, insuitable dosage unit formulations containing conventional non-toxicpharmaceutically acceptable carriers, adjuvants and vehicles appropriatefor each route of administration. The present disclosure alsocontemplates administration of the compounds and compositions of thepresent disclosure in a depot formulation.

In the treatment or prevention of conditions which require chemokinereceptor modulation an appropriate dosage level will generally be about0.001 to 100 mg per kg patient body weight per day which can beadministered in single or multiple doses. Preferably, the dosage levelwill be about 0.01 to about 25 mg/kg per day; more preferably about 0.05to about 10 mg/kg per day. A suitable dosage level may be about 0.01 to25 mg/kg per day, about 0.05 to 10 mg/kg per day, or about 0.1 to 5mg/kg per day. Within this range the dosage may be 0.005 to 0.05, 0.05to 0.5, 0.5 to 5.0, or 5.0 to 50 mg/kg per day. For oral administration,the compositions are preferably provided in the form of tabletscontaining 1.0 to 1000 milligrams of the active ingredient, particularly1.0, 5.0, 10.0, 15.0, 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0,250.0, 300.0, 400.0, 500.0, 600.0, 750.0, 800.0, 900.0, and 1000.0milligrams of the active ingredient for the symptomatic adjustment ofthe dosage to the patient to be treated. The compounds may beadministered on a regimen of 1 to 4 times per day, preferably once ortwice per day.

It will be understood, however, that the specific dose level andfrequency of dosage for any particular patient may be varied and willdepend upon a variety of factors including the activity of the specificcompound employed, the metabolic stability and length of action of thatcompound, the age, body weight, hereditary characteristics, generalhealth, sex, diet, mode and time of administration, rate of excretion,drug combination, the severity of the particular condition, and the hostundergoing therapy.

In some embodiments, compounds of the present disclosure areadministered as part of a combination therapy. For instance an amount ofa chemotherapeutic agent or radiation is administered to the subjectprior to, subsequent to or in combination with the compounds of thepresent disclosure. In some embodiments, the amount is sub-therapeuticwhen the chemotherapeutic agent or radiation is administered alone.Those of skill in the art will appreciate that “combinations” caninvolve combinations in treatments (i.e., two or more drugs can beadministered as a mixture, or at least concurrently or at leastintroduced into a subject at different times but such that both are inthe bloodstream of a subject at the same time). Additionally,compositions of the current disclosure may be administered prior to orsubsequent to a second therapeutic regimen, for instance prior to orsubsequent to a dose of chemotherapy or irradiation.

In still other embodiments, the present methods are directed to thetreatment of allergic diseases, wherein a compound or composition of thedisclosure is administered either alone or in combination with a secondtherapeutic agent, wherein said second therapeutic agent is anantihistamine or an anti-inflammatory. When used in combination, thepractitioner can administer a combination of the compound or compositionof the present disclosure and a second therapeutic agent. Also, thecompound or composition and the second therapeutic agent can beadministered sequentially, in any order.

The compounds and compositions of the present disclosure can be combinedwith other compounds and compositions having related utilities toprevent and treat the condition or disease of interest, such asinflammatory conditions and diseases, including inflammatory boweldisease (including Crohn's disease and ulcerative colitis), allergicdiseases, psoriasis, atopic dermatitis and asthma, and those pathologiesnoted above. Selection of the appropriate agents for use in combinationtherapies can be made one of ordinary skill in the art. The combinationof therapeutic agents may act synergistically to effect the treatment orprevention of the various disorders. Using this approach, one may beable to achieve therapeutic efficacy with lower dosages of each agent,thus reducing the potential for adverse side effects.

In treating, preventing, ameliorating, controlling or reducing the riskof inflammation, the compounds of the present disclosure may be used inconjunction with an antiinflammatory or analgesic agent such as anopiate agonist, a lipoxygenase inhibitor, such as an inhibitor of5-lipoxygenase, a cyclooxygenase inhibitor, such as a cyclooxygenase-2inhibitor, an interleukin inhibitor, such as an interleukin-1 inhibitor,an NMDA antagonist, an inhibitor of nitric oxide or an inhibitor of thesynthesis of nitric oxide, aminosalicylates, corticosteroids and otherimmunosuppressive drugs, a non-steroidal antiinflammatory agent, or acytokine-suppressing antiinflammatory agent, for example with a compoundsuch as acetaminophen, aspirin, codeine, biological TNF sequestrants,biological agents which target α4β7, ACE2 inhibitors, protein linase Cinhibitors, fentanyl, ibuprofen, indomethacin, ketorolac, morphine,naproxen, phenacetin, piroxicam, a steroidal analgesic, sufentanyl,sunlindac, tenidap, and the like.

Similarly, the compounds of the present disclosure may be administeredwith a pain reliever; a potentiator such as caffeine, an H2-antagonist,simethicone, aluminum or magnesium hydroxide; a decongestant such aspseudophedrine; an antitussive such as codeine; a diuretic; a sedatingor non-sedating antihistamine; a very late antigen (VLA-4) antagonist;an immunosuppressant such as cyclosporin, tacrolimus, rapamycin, EDGreceptor agonists, or other FK-506 type immunosuppressants; a steroid; anon-steroidal anti-asthmatic agent such as a β2-agonist, leukotrieneantagonist, or leukotriene biosynthesis inhibitor; an inhibitor ofphosphodiesterase type IV (PDE-IV); a cholesterol lowering agent such asa HMG-CoA reductase inhibitor, sequestrant, or cholesterol absorptioninhibitor; and an anti-diabetic agent such as insulin, α-glucosidaseinhibitors or glitazones.

The weight ratio of the compound of the present disclosure to the secondactive ingredient may be varied and will depend upon the effective doseof each ingredient. Generally, an effective dose of each will be used.Thus, for example, when a compound of the present disclosure is combinedwith an NSAID the weight ratio of the compound of the present disclosureto the NSAID will generally range from about 1000:1 to about 1:1000,preferably about 200:1 to about 1:200. Combinations of a compound of thepresent disclosure and other active ingredients will generally also bewithin the aforementioned range, but in each case, an effective dose ofeach active ingredient should be used.

Methods of Treating or Preventing CCR(9)-Mediated Conditions or Diseases

In yet another aspect, the present disclosure provides methods oftreating or preventing a CCR(9)-mediated condition or disease byadministering to a subject having such a condition or disease atherapeutically effective amount of any compound of formula I, I′, Ia,Ib, Ia1, Ia2, Ia3, Ia1′, Ia2′, Ia3′, Ib1, Ib2, Ib3, Ib1 Ib2′ or Ib3′.Compounds for use in the present methods include those compoundsaccording to the formula I, I′, Ia, Ib, Ia1, Ia2, Ia3, Ia1′, Ia2′, Ia3′,Ib1, Ib2, Ib3, Ib1′, Ib2′ and Ib3′, those provided above as embodiments,those specifically exemplified in the Examples below, and those providedwith specific structures herein. The “subject” is defined herein toinclude animals such as mammals, including, but not limited to, primates(e.g., humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats,mice and the like. In preferred embodiments, the subject is a human.

As used herein, the phrase “CCR(9)-mediated condition or disease” andrelated phrases and terms refer to a condition or disease characterizedby inappropriate, i.e., less than or greater than normal, CCR(9)functional activity. Inappropriate CCR(9) functional activity mightarise as the result of CCR(9) expression in cells which normally do notexpress CCR(9), increased CCR(9) expression (leading to, e.g.,inflammatory and immunoregulatory disorders and diseases) or decreasedCCR(9) expression. Inappropriate CCR(9) functional activity might alsoarise as the result of TECK secretion by cells which normally do notsecrete TECK, increased TECK expression (leading to, e.g., inflammatoryand immunoregulatory disorders and diseases) or decreased TECKexpression. A CCR(9)-mediated condition or disease may be completely orpartially mediated by inappropriate CCR(9) functional activity. However,a CCR(9)-mediated condition or disease is one in which modulation ofCCR(9) results in some effect on the underlying condition or disease(e.g., a CCR(9) antagonist results in some improvement in patientwell-being in at least some patients).

The term “therapeutically effective amount” means the amount of thesubject compound that will elicit the biological or medical response ofa cell, tissue, system, or animal, such as a human, that is being soughtby the researcher, veterinarian, medical doctor or other treatmentprovider.

Diseases and conditions associated with inflammation, immune disorders,infection and cancer may be treated or prevented with the presentcompounds, compositions, and methods. In one group of embodiments,diseases or conditions, including chronic diseases, of humans or otherspecies maybe treated with inhibitors of CCR(9) function. These diseasesor conditions include: (1) allergic diseases such as systemicanaphylaxis or hypersensitivity responses, drug allergies, insect stingallergies and food allergies, (2) inflammatory bowel diseases, such asCrohn's disease, ulcerative colitis, microscopic colitis, ileitis andenteritis, and postoperative ileus, (3) vaginitis, (4) psoriasis andinflammatory dermatoses such as dermatitis, eczema, atopic dermatitis,allergic contact dermatitis, urticaria and pruritus, (5) vasculitis, (6)spondyloarthropathies, (7) scleroderma, (8) asthma and respiratoryallergic diseases such as allergic asthma, allergic rhinitis,hypersensitivity lung diseases and the like, (9) autoimmune diseases,such as fibromyalagia, ankylosing spondylitis, juvenile RA, Still'sdisease, polyarticular juvenile RA, pauciarticular juvenile RA,polymyalgia rheumatica, rheumatoid arthritis, psoriatic arthritis,osteoarthritis, polyarticular arthritis, multiple sclerosis, systemiclupus erythematosus, type I diabetes, type II diabetes,glomerulonephritis, and the like, (10) graft rejection (includingallograft rejection), (11) graft-v-host disease (including both acuteand chronic), (12) other diseases in which undesired inflammatoryresponses are to be inhibited, such as atherosclerosis, myositis,neurodegenerative diseases (e.g., Alzheimer's disease), encephalitis,meningitis, hepatitis, nephritis, sepsis, sarcoidosis, allergicconjunctivitis, otitis, chronic obstructive pulmonary disease,sinusitis, Behcet's syndrome and gout, (13) immune mediated foodallergies such as Coeliac (Celiac) disease (14) pulmonary fibrosis andother fibrotic diseases, (15) irritable bowel syndrome, (16) primarysclerosing cholangitis, (17) cancer (including both primary andmetastatic), (18) bacterial associated syndromes such as hemorrhagiccolitis and hemolytic uremic syndrome (19) melanoma, (20) primarysclerosing cholangitis, (21) post-operative ileus (22) hepatitis andinflammatory hepatic diseases (23) Sjogren syndrome.

In another group of embodiments, diseases or conditions may be treatedwith modulators and agonists of CCR(9) function. Examples of diseasesthat may be treated by modulating CCR(9) function include cancers,cardiovascular diseases, diseases in which angiogenesis orneovascularization play a role (neoplastic diseases, retinopathy andmacular degeneration), infectious diseases (viral infections, e.g., HIVinfection, and bacterial infections) and immunosuppressive diseases suchas organ transplant conditions and skin transplant conditions. The term“organ transplant conditions” is means to include bone marrow transplantconditions and solid organ (e.g., kidney, liver, lung, heart, pancreasor combination thereof) transplant conditions.

Preferably, the present methods are directed to the treatment ofdiseases or conditions selected from inflammatory bowel diseaseincluding Crohn's disease and Ulcerative Colitis, allergic diseases,psoriasis, atopic dermatitis and asthma, autoimmune disease such asrheumatoid arthritis and immune-mediated food allergies such as Celiacdisease.

In yet other embodiments, the present methods are directed to thetreatment of psoriasis where a compound or composition of the disclosureis used alone or in combination with a second therapeutic agent such asa corticosteroid, a lubricant, a keratolytic agent, a vitamin D3derivative, PUVA and anthralin.

In other embodiments, the present methods are directed to the treatmentof atopic dermatitis using a compound or composition of the disclosureeither alone or in combination with a second therapeutic agent such as alubricant and a corticosteroid.

In further embodiments, the present methods are directed to thetreatment of asthma using a compound or composition of the disclosureeither alone or in combination with a second therapeutic agent such as a132-agonist and a corticosteroid.

Kits and Packages

The terms “kit” and “pharmaceutical kit” refer to a commercial kit orpackage comprising, in one or more suitable containers, one or morepharmaceutical compositions and instructions for their use. In oneembodiment, kits comprising a compound of Formula I, I′, Ia, Ib, Ia1,Ia2, Ia3, Ia1′, Ia2′, Ia3′, Ib1, Ib2, Ib3, Ib1′, Ib2′ or Ib3′, or apharmaceutically acceptable salt thereof, and instructions for itsadministration are provided. In one embodiment, kits comprising acompound of Formula I, I′, Ia, Ib, Ia1, Ia2, Ia3, Ia1′, Ia2′, Ia3′, Ib1,Ib2, Ib3, Ib1′, Ib2′ or Ib3′, or a pharmaceutically acceptable saltthereof, in combination with one or more (e.g., one, two, three, one ortwo, or one to three) additional therapeutic agents and instructions fortheir administration are provided.

In one embodiment, the compounds of this disclosure are formulated intoadministration units which are packaged in a single packaging. Thesingle packaging encompasses but is not limited to a bottle, achild-resistant bottle, an ampoule, and a tube. In one embodiment, thecompounds of this disclosure and optionally additional therapeuticagents, are formulated into administration units and every singleadministration unit is individually packaged in a single packaging. Suchindividually packaged units may contain the pharmaceutical compositionin any form including but not limited to liquid form, solid form, powderform, granulate form, an effervescent powder or tablet, hard or softcapsules, emulsions, suspensions, syrup, suppositories, tablet, troches,lozenges, solution, buccal patch, thin film, oral gel, chewable tablet,chewing gum, and single-use syringes. Such individually packaged unitsmay be combined in a package made of one or more of paper, cardboard,paperboard, metal foil and plastic foil, for example a blister pack. Oneor more administration units may be administered once or several times aday. One or more administration units may be administered three times aday. One or more administration units may be administered twice a day.One or more administration units may be administered on a first day andone or more administration units may be administered on the followingdays.

Additional Combination Therapies

The compounds of this disclosure can be supplied alone or in conjunctionwith one or more other drugs. Examples of therapeutic agents that may becombined with a compound or composition of the present disclosure,either administered separately or in the same pharmaceuticalcomposition, include, but are not limited to: modulators of CCR1, CCR2,CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CCR10, CCR11, CXCR1, CXCR2,CXCR3, CXCR4, CXCR5, CXCR6, CXCR7, CX3CR1, ChemR23, C5aR, C5a, and C5,or any combination thereof. In some embodiments, the modulator is anantagonist.

Examples of therapeutic agents that may be combined with a compound orcomposition of the present disclosure, either administered separately orin the same pharmaceutical composition, include, but are not limited to:CCX354, CCX9588, CCX140, CCX872, CCX598, CCX6239, CCX9664, CCX2553, CCX2991, CCX282, CCX025, CCX507, CCX430, CCX765, CCX224, CCX662, CCX650,CCX832, CCX168, CCX168-M1, brazikumab, budesonide, ustekinumab,everolimus, glatiramer acetate, natalizumab, etanercept, mycophenolatemofetil, brodalumab, cannabidiol, foralumab, tralokinumab, tamibarotene,mesalazine, golimumab, teduglutide, infliximab, ropivacaine, filgotinib,etrolizumab, SHP-647, elafibranor, ABC-294640, ocrelizumab, tofacitinib,certolizumab pegol, adalimumab, sargramostim, abatacept, clarithromycin,GSK-2982772, upadacitinib, edasalonexent, secukinumab, vancomycin,vedolizumab, thalidomide, rituximab, catridecacog, RBX-2660, Ampion,nitazoxanide, fingolimod, tocilizumab, rosiptor acetate, AST-120,risankizumab, telotristat etiprate, lenalidomide, alicaforsen,tosufloxacin, interferon beta-1a, E-6011, KAG-308, dexamethasone sodiumphosphate, ozanimod, dociparstat cobitolimod, mesalazine, PUR-0110,apremilast, mesalazine, valganciclovir, tacrolimus, mongersen,remestemcel-L, GS-5745, E-6011, E-6007, carotegrast methyl,piclidenoson, PF-06480605, balsalazide, pimecrolimus, mesalazine,recombinant interferon beta-1a, naltrexone, adalimumab, amiselimod,brilacidin, basiliximab, etrasimod, LP-02, rosiglitazone, plecanatide,laquinimod, rifabutin+clarithromycin+clofazimine, infliximab,tildrakizumab, omega-3-carboxylic acids, TOP-1288, peficitinib,rifamycin, rifaximin, JNJ-64304500, ASP-3291, DLX-105, zileuton, 99mTclabelled annexin V-128, ALT-836, Biferonex, clotrimazole, givinostat,Trichuris suis ova, INV-103, K(D)PT, BI-655064, glepaglutide, LYC-30937EC, TRX-318, LY-3074828, larazotide acetate, IBP-9414, clazakizumab,mesalazine, eclomethasone dipropionate, NN-8828, olokizumab,bertilimumab, midismase, KRP-203, prednisolone, PF-06687234, STNM-01,KHK-4083, FE-999301, DLX-105, VB-201, DNVX-078, rifaximin, Clostridiumbutyricum MIYAIRI 588, OPS-2071, sotrastaurin, abrilumab, QBECO,anakinra, FFP-104, GLPG-1205, dolcanatide, PDA-002, molgramostim,mesalazine, metronidazole, repurposed naltrexone, vatelizumab,zucapsaicin, ciclosporin, oprelvekin, prulifloxacin, recombinant humanlactoferrin, Alequel, SAN-300, STP-206, GLPG-0974, P-28-GST, N-6022, TNFalpha kinoid, ETX-201, low molecular weight heparin, ETX-201,GED-0507-34-Levo, metenkefalin acetate+tridecactide acetate, HMPL-004,SB-012, TRK-170, beta-1,3/1,6-glucan, mesalamine+N-acetylcysteine,99mTc-sulesomab, olsalazine, mesalazine Bacillus licheniformis,balsalazide sodium, propionyl-L-carnitine, Clostridium butyricum,beclomethasone dipropionate, acemannan, and SPD-480, or any combinationthereof.

Examples of therapeutic agents that may be combined with a compound orcomposition of the present disclosure, either administered separately orin the same pharmaceutical composition, include, but are not limited to:an IL-23 antagonist, a Glucocorticoid agonist, an IL-6 agonist, an IL-12antagonist, a mTOR complex 1 inhibitor, a mTOR inhibitor, a celladhesion molecule inhibitor, an Integrin alpha-4/beta-1 antagonist, aTNF antagonist, a TNF binding agent, a Type II TNF receptor modulator,an Inosine monophosphate dehydrogenase inhibitor, a PurH purinebiosynthesis protein inhibitor, an Interleukin receptor 17A, antagonist,a Cannabinoid CB1 receptor modulator, a Cannabinoid CB2 receptormodulator, a Cannabinoid receptor modulator, a CD3 antagonist, an IL-13antagonist, a Retinoic acid receptor alpha agonist, a Retinoic acidreceptor beta agonist, a Retinoid receptor agonist, a Cyclooxygenaseinhibitor, a TNF alpha ligand inhibitor, a Glucagon-like peptide 2agonist, a sodium channel inhibitor, a Jak1 tyrosine kinase inhibitor,an Integrin alpha-4/beta-7 antagonist, an Integrin alpha-E antagonist,an Integrin beta-7 antagonist, an Immunoglobulin G2 modulator, a MAdCAMinhibitor, an Insulin sensitizer, a PPAR alpha agonist, a PPAR deltaagonist, a Collagen modulator, a Dihydroceramide delta 4 desaturaseinhibitor, a Sphingosine kinase 1 inhibitor, a Sphingosine kinase 2inhibitor, aB-lymphocyte antigen CD20 inhibitor, a JAK tyrosine kinaseinhibitor, a Jak3 tyrosine kinase inhibitor, a CSF-1 agonist, a GM-CSFreceptor agonist, a Cytotoxic T-lymphocyte protein-4 stimulator, a Tcell surface glycoprotein CD28 inhibitor, a RIP-1 kinase inhibitor, aNuclear factor kappa B inhibitor, an IL-17 antagonist, a Peptidoglycanrecognition protein inhibitor, an Integrin alpha-4/beta-7 antagonist, aB-lymphocyte antigen CD20 inhibitor, a Factor XIII agonist, a Stem cellantigen-1 inhibitor, a Cannabinoid receptor antagonist;Sphingosine-1-phosphate receptor-1 modulator, an IL-6 antagonist, anIL-6 receptor modulator, a SH2 domain inositol phosphatase 1 stimulator,a Tryptophan 5-hydroxylase inhibitor, a ICAM1 gene inhibitor, a DNAgyrase inhibitor, a Topoisomerase IV inhibitor, an Interferon betaligand, a Fractalkine ligand inhibitor, a EP4 prostanoid receptoragonist, a Sphingosine-1-phosphate receptor-1 agonist, a Sphingosinephosphate receptor-1 modulator, a Sphingosine-1-phosphate receptor-5modulator, a Cathepsin G inhibitor, a Complement cascade inhibitor, anElastase inhibitor, a Heparin agonist, a L-Selectin antagonist, aP-Selectin antagonist, a Nuclear factor kappa B inhibitor, a TLR-9agonist, an Interleukin-1 beta ligand modulator, a PDE 4 inhibitor, aDNA polymerase inhibitor, a SMAD-7 inhibitor, a TGF beta 1 ligandinhibitor, a Metalloprotease-9 inhibitor, a Fractalkine ligandinhibitor, an Integrin antagonist, an Adenosine A3 receptor agonist, aTumor necrosis factor 15 ligand inhibitor, an IL-10 antagonist, an IL-2antagonist, an IL-4 antagonist, an Interferon gamma receptor antagonist,an Interferon beta ligand, an Opioid receptor antagonist, an IL-2receptor alpha subunit inhibitor, a Sphingosine 1 phosphate phosphatase1 stimulator, an Insulin sensitizer, a PPAR gamma agonist, a Natriureticpeptide receptor C agonist, a n acyltransferase inhibitor, anapolipoprotein C3 antagonist, an adapter molecule crk inhibitor, an IL-8antagonist, an Interleukin-1 beta ligand inhibitor, a Src tyrosinekinase inhibitor, a Syk tyrosine kinase inhibitor, a DNA RNA polymeraseinhibitor, a RNA polymerase inhibitor, a Melanocortin agonist, a5-Lipoxygenase inhibitor, a Tissue factor inhibitor, an Interferon betaligand, a Bradykinin receptor modulator, an Histone deacetylaseinhibitor, a P2X7 purinoceptor agonist, a mitochondrial 10 kDa heatshock protein stimulator, a CD40 ligand receptor antagonist, aGlucagon-like peptide 2 agonist, a F1F0 ATP synthase modulator, a CD3antagonist, a Zonulin inhibitor, a Cyclooxygenase inhibitor, aLipoxygenase modulator, an IL-21 antagonist, a CCR3 chemokineantagonist, an Eotaxin ligand inhibitor, a Superoxide dismutasemodulator, a Sphingosine-1-phosphate receptor-1 agonist, a CD29modulator, an Interleukin-10 ligand, a CHST15 gene inhibitor, an OX40ligand inhibitor, an IL-6 receptor modulator, a Nuclear factor kappa Binhibitor, an Oncostatin M receptor modulator, a STAT inhibitor, aSTAT-3 inhibitor, a TLR-2 antagonist, a TLR-4 antagonist, a RNApolymerase inhibitor, a Protein kinase C alpha inhibitor, a Proteinkinase C beta inhibitor, a Protein kinase C delta inhibitor, a Proteinkinase C epsilon inhibitor, a Protein kinase C eta inhibitor, a Proteinkinase C theta inhibitor, a Type I IL-1 receptor antagonist, a CD40ligand inhibitor, a CD40 ligand receptor antagonist, a G-protein coupledreceptor 84 antagonist, a Guanylate cyclase receptor agonist, a CD49bantagonist, a Vanilloid VR1 agonist, a Calcineurin inhibitor, an IL-11agonist, a PDGF receptor agonist, a DNA gyrase inhibitor, a Lactoferrinstimulator, an Integrin alpha-1/beta-1 antagonist, a Free fatty acidreceptor 2 antagonist, an Alcohol dehydrogenase 5 inhibitor, aglutathione reductase inhibitor, an Interferon gamma receptorantagonist, Low molecular weight heparin, a PPAR gamma agonist, a ACTHreceptor agonist, an Adrenocorticotrophic hormone ligand, an Opioidgrowth factor receptor agonist, an IL-6 antagonist, an Interleukin-1beta ligand modulator, a Nuclear factor kappa B inhibitor; a GATA 3transcription factor inhibitor, a Nuclear factor kappa B inhibitor, anOxidoreductase inhibitor, a Glucocorticoid agonist, an Interferon gammareceptor agonist, or any combination thereof.

EXAMPLES

The following examples are offered to illustrate, but not to limit theclaimed disclosure.

Reagents and solvents used below can be obtained from commercial sourcessuch as Aldrich Chemical Co. (Milwaukee, Wis., USA). ¹H-NMR wererecorded on a Varian Mercury 400 MHz NMR spectrometer. Significant peaksare provided relative to TMS and are tabulated in the order:multiplicity (s, singlet; d, doublet; t, triplet; q, quartet; m,multiplet) and number of protons. Mass spectrometry results are reportedas the ratio of mass over charge, followed by the relative abundance ofeach ion (in parenthesis). In tables, a single m/e value is reported forthe M+H (or, as noted, M−H) ion containing the most common atomicisotopes. Isotope patterns correspond to the expected formula in allcases. Electrospray ionization (ESI) mass spectrometry analysis wasconducted on a Hewlett-Packard MSD electrospray mass spectrometer usingthe HP1100 HPLC equipped with an Agilent Zorbax SB-C18, 2.1×50 mm, 5μcolumn for sample delivery. Normally the analyte was dissolved inmethanol at 0.1 mg/mL and 1 microlitre was infused with the deliverysolvent into the mass spectrometer, which scanned from 100 to 1500daltons. All compounds could be analyzed in the positive ESI mode, usingacetonitrile/water with 1% formic acid as the delivery solvent. Thecompounds provided below could also be analyzed in the negative ESImode, using 2 mM NH₄OAc in acetonitrile/water as delivery system.

The following abbreviations are used in the Examples and throughout thedescription of the disclosure:

-   -   HPLC, High Pressure Liquid Chromatography; DMF, Dimethyl        formamide; TFA, Trifluoroacetic Acid; THF, Tetrahydrofuran;        EtOAc, Ethyl acetate; BOC₂O, di-tertbutyl dicarbonate or BOC        anhydride; HPLC, High Pressure Liquid Chromatography; DIPEA,        Diisopropyl ethylamine; HBTU,        O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium        hexafluorophosphate; dppf, 1,1′-Bis(diphenylphosphino)ferrocene;        Pd₂(dba)₃, Tris(dibenzylideneacetone)dipalladium(0); DIPEA,        diisopropylethylamine; DMP, dimethylphthalate; Me, methyl; Et,        ethyl; DCM, dichloromethane.

Compounds within the scope of this disclosure can be synthesized asdescribed below, using a variety of reactions known to the skilledartisan. One skilled in the art will also recognize that alternativemethods may be employed to synthesize the target compounds of thisdisclosure, and that the approaches described within the body of thisdocument are not exhaustive, but do provide broadly applicable andpractical routes to compounds of interest.

Certain molecules disclosed in this patent can exist in differentenantiomeric and diastereomeric forms and all such variants of thesecompounds are claimed.

The detailed description of the experimental procedures used tosynthesize key compounds in this text lead to molecules that aredescribed by the physical data identifying them as well as by thestructural depictions associated with them.

Those skilled in the art will also recognize that during standard workup procedures in organic chemistry, acids and bases are frequently used.Salts of the parent compounds are sometimes produced, if they possessthe necessary intrinsic acidity or basicity, during the experimentalprocedures described within this patent.

Example 1: Synthesis of(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-indolin-2-one

a) A solution of silver triflate (0.385 g, 1.50 mmol) in1,2-dichloroethane (1.0 L) was heated at 90° C. for 3 h under a refluxcondenser and nitrogen atmosphere. The cloudy mixture was cooled to roomtemperature before methyl 4-hydroxyphenylacetate (24.9 g, 150 mmol) wasadded. Isoprene (15.3 g, 225 mmol) in 1,2-dichloroethane (100 mL) wasadded dropwise over 10 min and the mixture was stirred at roomtemperature for 2 h. The solution was concentrated, diluted in EtOAc(150 mL), and washed with saturated aqueous NaHCO₃ (2×100 mL) and 1 MNaHSO₄ (2×100 mL). The organic layer was dried over Na₂SO₄, filtered,and concentrated in vacuo. Purification of the crude material by flashchromatography (1-10% EtOAc in hexanes) eluted methyl2-(2,2-dimethylchroman-6-yl)acetate.

b) To a cooled (−30° C.) solution of sodium tert-butoxide (93.0 g, 963mmol) in anhydrous N-methylpyrrolidone (500 mL) under nitrogenatmosphere was slowly added methyl 2-(2,2-dimethylchroman-6-yl)acetate(80.8 g, 344 mmol), followed by 4-chloro fluoronitrobenzene (62.1 g, 354mmol), and more N-methylpyrrolidone (150 mL). After stirring at −30° C.for 1 h, methyl iodide (42.8 mL, 688 mmol) was added by syringe and themixture was stirred for an additional 20 min. The reaction was quenchedby addition of 3 M HCl (600 mL) and the mixture was warmed to roomtemperature. The reaction mixture was extracted with methyl tert-butylether (1×500 mL), dried over Na₂SO₄, filtered through a plug of silicagel, and concentrated in vacuo to afford a brown solid. The crudematerial was diluted in a solution of isopropyl alcohol (520 mL), water(260 mL), and ethylene glycol (340 g) and then cooled to 0° C. Potassiumhydroxide (130 g, 2.32 mol) was added and the solution was heated to100° C. for 4 h. After the completion of the reaction, the mixture wascooled back to 0° C., acidified to pH 3 with aqueous 3 M HCl, andextracted with EtOAc (2×300 mL). The organic layers were dried overNa₂SO₄, filtered through a plug of silica gel, and concentrated in vacuoto obtain2-(5-chloro-2-nitro-phenyl)-2-(2,2-dimethylchroman-6-yl)propionic acid.

c) Oxalyl chloride (2.10 mL, 24.0 mmol) and dimethylformamide (0.10 mL)were sequentially added to a stirred solution of2-(5-chloro-2-nitro-phenyl)-2-(2,2-dimethylchroman-6-yl)propionic acid(7.21 g, 18.5 mmol) in dichloromethane (100 mL) at room temperature.After 1.5 h, the reaction mixture was concentrated and re-dissolved indichloromethane (100 mL). Triethylamine (7.77 mL, 55.2 mmol) and(S)-(+)-phenylglycinol (2.54 g, 18.5 mmol) were added and the mixturewas allowed to stir at room temperature until the reaction was complete(1 h). The mixture was concentrated, diluted in EtOAc (300 mL), andwashed with aqueous 1 M HCl (1×200 mL). The organic layer was dried overNa₂SO₄, filtered, and concentrated in vacuo. Purification of the crudematerial by flash chromatography (1-8% THF in DCM) separated thediastereomers of2-(5-chloro-2-nitro-phenyl)-2-(2,2-dimethylchroman-6-yl)-N-(2-hydroxy-1-phenyl-ethyl)propanamide.The second-eluting diastereomer was taken through subsequenttransformations.

d) To a solution of2-(5-chloro-2-nitro-phenyl)-2-(2,2-dimethylchroman-6-yl)-N-(2-hydroxy-1-phenyl-ethyl)propanamide(3.95 g, 7.76 mmol) and acetic acid (4 mL) in methanol (80 mL) was addediron powder (4.0 g, 71.7 mmol) and the reaction mixture was heated to70° C. for 2 h. After cooling to room temperature, the mixture wasdiluted with EtOAc (150 mL) and washed with 1 M HCl (1×100 mL) and water(1×100 mL). The organic layer was dried over Na₂SO₄, filtered, andconcentrated in vacuo. Purification of the crude material by flashchromatography (0-40% EtOAc in hexanes) gave(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-indolin-2-one.

Example 2: Alternate Synthesis of(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-indolin-2-one

A solution of2-(5-chloro-2-nitro-phenyl)-2-(2,2-dimethylchroman-6-yl)propionic acid(37.3 g, 95.6 mmol) and (S)-1-(4-chlorophenyl)ethylamine in isopropylalcohol (150 mL) and water (50 mL) was heated at 100° C. until allsolids were dissolved. The solution was then allowed to gradually coolto room temperature and left to sit undisturbed overnight. The salt thatcame out of solution was filtered and washed with 2:1 IPA-H₂O (180 mL)to give a pure crystalline material (17.0 g, 33%, er>100:1 as the freefrom). A solution of crystalline (S)-1-(4-chlorophenyl)ethylamine saltof 2-(5-chloro-2-nitro-phenyl)-2-(2,2-dimethylchroman-6-yl)propionicacid (546 mg, 1.0 mmol), iron powder (224 mg, 4.0 mmol), and acetic acid(480 mg, 8.0 mmol) in methanol (5.0 mL) was heated to 70° C. for 1 h.After cooling to room temperature, the mixture was diluted with EtOAc(50 mL) and washed with 1 M HCl (1×50 mL) and water (1×50 mL). Theorganic layer was dried over Na₂SO₄, filtered, and concentrated invacuo. Purification by flash chromatography (0-40% EtOAc in hexanes)provided(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-indolin-2-one.

Example 3: Synthesis of3-[(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-2-oxo-indolin-1-yl]benzoicacid

To a solution of(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-indolin-2-one (45mg, 0.13 mmol), tert-butyl-3-iodobenzoate (80 mg, 0.27 mmol),trans-N,N′-dimethylcyclohexane-1,2-diamine (10 mg, 0.07 mmol), andpotassium carbonate (70 mg, 0.51 mmol) in dioxane (2.5 mL) was addedcopper iodide (10 mg, 0.053 mmol). The mixture was purged with nitrogenand heated to 100° C. After 1 h, the mixture was cooled to roomtemperature and diluted with EtOAc (20 mL). The organic layer was washedwith 1 M HCl (1×20 mL), water (1×20 mL), dried over Na₂SO₄, filtered,and concentrated in vacuo. The crude material was diluted indichloromethane (1 mL) and trifluoroacetic acid (2 mL) and stirred atroom temperature for 5 h. The mixture was concentrated and purified byreverse-phase HPLC (C18 column, acetonitrile-H₂O with 0.1% TFA aseluent) to afford the titled compound. ¹H NMR (400 MHz, Chloroform-d) δ8.22-8.09 (m, 2H), 7.69 (dddd, J=8.0, 1.8, 1.3, 0.4 Hz, 1H), 7.66-7.58(m, 1H), 7.26-7.17 (m, 2H), 7.06 (d, J=2.3 Hz, 1H), 6.99 (ddd, J=8.5,2.5, 0.6 Hz, 1H), 6.88-6.81 (m, 1H), 6.73 (d, J=8.6 Hz, 1H), 2.76 (t,J=6.7 Hz, 2H), 1.87 (s, 3H), 1.78 (t, J=6.7 Hz, 2H), 1.32 (d, J=1.5 Hz,6H);); MS: (ES) m/z calculated for C₂₇H₂₅ClNO₄ [M+H]⁺ 462.1, found462.5.

Example 4: Synthesis of4-[[(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-2-oxo-indolin-1-yl]methyl]benzoicacid

To a cooled (0° C.) solution of(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-indolin-2-one (45mg, 0.13 mmol) in anhydrous dimethylformamide (0.80 mL) under nitrogenwas added sodium hydride (20 mg, 60% suspension in mineral oil, 0.50mmol). After stirring at 0° C. for 10 min, the solution was allowed towarm to room temperature and methyl 4-(chloromethyl)benzoate (25 mg,0.14 mmol) was added. The mixture was left to stir for 30 min at roomtemperature before the reaction was carefully quenched by the additionof 1 M HCl (25 mL) and extracted with EtOAc (50 mL). The organic layerwas washed with water (1×20 mL), dried over Na₂SO₄, filtered, andconcentrated in vacuo. The crude material was re-dissolved in a mixtureof ethanol (2 mL) and water (1 mL). Lithium hydroxide monohydrate (100mg, 2.4 mmol) was added and the mixture was stirred at 50° C. for 30min. After cooling to room temperature, the reaction was quenched by theaddition of 1 M HCl (25 mL) and extracted with EtOAc (50 mL). Theorganic layer was washed with water (1×20 mL), dried over Na₂SO₄,filtered, and concentrated in vacuo. Purification of the crude materialby reverse-phase HPLC (C18 column, acetonitrile-H₂O with 0.1% TFA aseluent) gave the titled compound. ¹H NMR (400 MHz, Chloroform-d) δ 8.05(d, J=8.2 Hz, 2H), 7.34 (d, J=8.1 Hz, 2H), 7.19-7.12 (m, 2H), 6.99 (d,J=2.4 Hz, 1H), 6.91 (dd, J=8.6, 2.5 Hz, 1H), 6.72 (d, J=8.5 Hz, 1H),6.67-6.60 (m, 1H), 5.09-4.89 (m, 2H), 2.74 (t, J=6.8 Hz, 2H), 1.81 (s,3H), 1.78 (t, J=6.7 Hz, 2H), 1.32 (s, 6H); MS: (ES) m/z calculated forC₂₈H₂₇ClNO₄ [M+H]⁺ 476.1, found 476.2.

Example 5: Synthesis of3-[(3R)-3-(4-tert-butylphenyl)-5-chloro-3-methyl-2-oxo-indolin-1-yl]benzoicacid

a) To a cooled (0° C.) solution of sodium hydride (1.30 g, 39.9 mmol) inanhydrous dimethylformamide (5 mL) under nitrogen atmosphere was slowlyadded methyl p-tert-butylphenylacetate (1.70 g, 8.22 mmol) indimethylformamide (5 mL) and the mixture was allowed to stir at 0° C.for 30 min. Next, 4-chloro-2-fluoronitrobenzene (1.60 g, 9.05 mmol) indimethylformamide (3 mL) was added dropwise over 10 min. After 1.5 h ofstirring at 0° C., methyl iodide (0.51 mL, 16.4 mmol) was added and themixture was allowed to warm to room temperature and stir for anadditional 3 h. The reaction was quenched by the addition of saturatedaqueous NH₄C₁ (50 mL). The mixture was extracted with EtOAc (2×50 mL)and the organic layers were dried over Na₂SO₄, filtered, andconcentrated in vacuo. Purification by flash chromatography (0-20% EtOAcin hexanes) provided methyl2-(4-tert-butylphenyl)-2-(3-chlorophenyl)propionate. The compound wasdissolved in isopropyl alcohol (40 mL) and water (20 mL) with potassiumhydroxide (2.3 g, 41.1 mmol) and the mixture was heated at 100° C. for 2h. The mixture was cooled to room temperature, acidified to pH 3 with 1M HCl, and extracted with EtOAc (2×50 mL). The combined organic layerswere dried over Na₂SO₄, filtered, and concentrated in vacuo.Purification of the crude material by flash chromatography (0-30% EtOAcin hexanes) provided 2-(4-tert-butylphenyl)-2-(3-chlorophenyl)propionicacid.

b) Oxalyl chloride (0.57 mL, 6.6 mmol) and dimethylformamide (4 drops)were sequentially added to a stirred solution of2-(4-tert-butylphenyl)-2-(3-chlorophenyl)propionic acid (2.11 g, 5.5mmol) in dichloromethane (35 mL) at room temperature. After 2 h, thereaction mixture was concentrated and re-dissolved in dichloromethane(30 mL). Triethylamine (2.3 mL, 16.5 mmol) and (S)-(+)-phenylglycinol(750 mg, 5.5 mmol) were added and the mixture was allowed to stir atroom temperature for 1 h. The mixture was concentrated, diluted in EtOAc(100 mL) and washed with aqueous 1 M HCl (1×50 mL). The organic layerwas dried over Na₂SO₄, filtered, and concentrated in vacuo. Purificationof the crude material by flash chromatography (1-15% EtOAc in DCM)separated the diastereomers of(2R)-2-(4-tert-butylphenyl)-2-(3-chlorophenyl)-N-[(1R)-2-hydroxy-1-phenyl-ethyl)]propanamide.The second-eluting diastereomer was taken through subsequenttransformations.

c) To a solution of(2R)-2-(4-tert-butylphenyl)-2-(3-chlorophenyl)-N-[(1R)-2-hydroxy-1-phenyl-ethyl)]propanamide(250 mg, 0.52 mmol) and acetic acid (0.22 mL) in methanol (2.6 mL) wasadded iron powder (87 mg, 1.56 mmol) and the reaction mixture was heatedto 70° C. for 2 h. After cooling to room temperature, the mixture wasdiluted with EtOAc (20 mL) and washed with 1 M HCl (1×10 mL) and water(1×10 mL). The organic layer was dried over Na₂SO₄, filtered, andconcentrated in vacuo. Purification of the crude material by flashchromatography (0-40% EtOAc in hexanes) gave(3R)-3-(4-tert-butylphenyl)-5-chloro-3-methyl-indolin-2-one.

d) To a solution of(3R)-3-(4-tert-butylphenyl)-5-chloro-3-methyl-indolin-2-one (34 mg, 0.11mmol), tert-butyl-3-iodobenzoate (65 mg, 0.22 mmol),trans-N,N′-dimethylcyclohexane-1,2-diamine (10 mg, 0.07 mmol), andpotassium carbonate (70 mg, 0.51 mmol) in dioxane (2.5 mL) was addedcopper iodide (10 mg, 0.053 mmol). The mixture was purged with nitrogenand heated to 100° C. After 1 h, the mixture was cooled to roomtemperature and diluted with EtOAc (20 mL). The organic layer was washedwith 1 M HCl (1×20 mL), water (1×20 mL), dried over Na₂SO₄, filtered,and concentrated in vacuo. The crude material was diluted indichloromethane (1 mL) and trifluoroacetic acid (2 mL) and stirred atroom temperature for 5 h. The mixture was concentrated and purified byreverse-phase HPLC (C18 column, acetonitrile-H₂O with 0.1% TFA aseluent) to afford the titled compound. ¹H NMR (400 MHz, Chloroform-d) δ8.18-8.10 (m, 2H), 7.72-7.59 (m, 2H), 7.40-7.35 (m, 2H), 7.29 (d, J=2.1Hz, 1H), 7.28-7.26 (m, 1H), 7.24 (d, J=7.7 Hz, 2H), 6.87-6.81 (m, 1H),1.91 (s, 3H), 1.30 (s, 9H); MS: (ES) m/z calculated for C₂₆H₂₅ClNO₃[M+H]⁺ 434.1, found 434.2.

Example 6: Synthesis of5-[(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-2-oxo-indolin-1-yl]-2-methoxy-benzoicacid

To a solution of(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methylindolin-2-one (60 mg,0.18 mmol), methyl 5-iodo-2-methoxybenzoate (80 mg, 0.27 mmol),trans-N,N′-dimethylcyclohexane-1,2-diamine (10 mg, 0.07 mmol), andpotassium carbonate (102 mg, 0.73 mmol)) in dioxane (3.0 mL) was addedcopper iodide (20 mg, 0.10 mmol). The mixture was purged with nitrogenand heated to 100° C. After 1 h, the mixture was cooled to roomtemperature and diluted with EtOAc (20 mL). The organic layer was washedwith 1 M HCl (1×20 mL), water (1×20 mL), dried over Na₂SO₄, filtered,and concentrated in vacuo. The crude material was re-dissolved in amixture of ethanol (2 mL) and water (1 mL). Lithium hydroxidemonohydrate (100 mg, 2.4 mmol) was added and the mixture was stirred at50° C. for 30 min. After cooling to room temperature, the reaction wasquenched by the addition of 1 M HCl (25 mL) and extracted with EtOAc (50mL). The organic layer was washed with water (1×20 mL), dried overNa₂SO₄, filtered, and concentrated in vacuo. Purification of the crudematerial by reverse-phase HPLC (C18 column, acetonitrile-H₂O with 0.1%TFA as eluent) gave the titled compound. ¹H NMR (400 MHz, Chloroform-d)δ 8.21 (d, J=2.7 Hz, 1H), 7.66 (dd, J=8.9, 2.7 Hz, 1H), 7.29-7.16 (m,3H), 7.01 (d, J=2.4 Hz, 1H), 6.96 (dd, J=8.6, 2.5 Hz, 1H), 6.78 (dd,J=8.1, 0.7 Hz, 1H), 6.73 (d, J=8.6 Hz, 1H), 4.13 (s, 3H), 2.75 (t, J=6.7Hz, 2H), 1.85 (s, 3H), 1.78 (t, J=6.7 Hz, 2H), 1.32 (s, 6H); MS: (ES)m/z calculated for C₂₈H₂₇ClNO₅ [M+H]⁺ 492.2, found 492.3.

Example 7: Synthesis of5-[(3R)-3-(2,2-dimethylchroman-6-yl)-3,5-dimethyl-2-oxo-indolin-1-yl]-2-methyl-benzoicacid

To a solution of(3R)-3-(2,2-dimethylchroman-6-yl)-3,5-dimethylindolin-2-one (32 mg, 0.10mmol), methyl 5-iodo-2-methylbenzoate (36 mg, 0.13 mmol),trans-N,N′-dimethylcyclohexane-1,2-diamine (10 mg, 0.07 mmol), andpotassium carbonate (28 mg, 0.20 mmol)) in dioxane (3.0 mL) was addedcopper iodide (6.0 mg, 0.03 mmol). The mixture was purged with nitrogenand heated to 100° C. After 1 h, the mixture was cooled to roomtemperature and diluted with EtOAc (20 mL). The organic layer was washedwith 1 M HCl (1×20 mL), water (1×20 mL), dried over Na₂SO₄, filtered,and concentrated in vacuo. The crude material was re-dissolved in amixture of ethanol (2 mL) and water (1 mL). Lithium hydroxidemonohydrate (100 mg, 2.4 mmol) was added and the mixture was stirred at50° C. for 30 min. After cooling to room temperature, the reaction wasquenched by the addition of 1 M HCl (25 mL) and extracted with EtOAc (50mL). The organic layer was washed with water (1×20 mL), dried overNa₂SO₄, filtered, and concentrated in vacuo. Purification of the crudematerial by reverse-phase HPLC (C18 column, acetonitrile-H₂O with 0.1%TFA as eluent) gave the titled compound. ¹H NMR (400 MHz, Chloroform-d)δ 8.19 (t, J=1.9 Hz, 1H), 8.13-8.06 (m, 1H), 7.76-7.69 (m, 1H), 7.61 (t,J=7.9 Hz, 1H), 7.14-7.03 (m, 3H), 7.03-6.97 (m, 1H), 6.86-6.77 (m, 1H),6.71 (d, J=8.6 Hz, 1H), 2.75 (t, J=6.8 Hz, 2H), 2.36 (d, J=0.9 Hz, 3H),1.85 (s, 3H), 1.78 (t, J=6.7 Hz, 2H), 1.31 (d, J=1.8 Hz, 6H); MS: (ES)m/z calculated for C₂₉H₃₀NO₄ [M+H]⁺ 456.6, found 456.0.

Example 8: Synthesis of2-[4-[(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyloxo-indolin-1-yl]phenyl]acetic acid

To a solution of(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-indolin-2-one (60mg, 0.18 mmol), 4-iodophenylacetic acid (92 mg, 0.35 mmol),trans-N,N′-dimethylcyclohexane-1,2-diamine (10 mg, 0.07 mmol), andpotassium carbonate (97 mg, 0.70 mmol) in dimethylformamide (3.0 mL) wasadded copper iodide (20 mg, 0.10 mmol). The mixture was purged withnitrogen and heated to 110° C. After 1 h, the mixture was cooled to roomtemperature and diluted with EtOAc (20 mL). The organic layer was washedwith 1 M HCl (1×20 mL), water (1×20 mL), dried over Na₂SO₄, filtered,and concentrated in vacuo. Purification of the crude material byreverse-phase HPLC (C18 column, acetonitrile-H₂O with 0.1% TFA aseluent) gave the titled compound. ¹H NMR (400 MHz, Chloroform-d) δ7.49-7.41 (m, 2H), 7.41-7.34 (m, 2H), 7.20 (dq, J=4.3, 2.1 Hz, 2H), 7.04(d, J=2.3 Hz, 1H), 6.96 (ddd, J=8.6, 1.9, 1.2 Hz, 1H), 6.85 (d, J=8.9Hz, 1H), 6.71 (d, J=8.5 Hz, 1H), 3.71 (s, 2H), 2.74 (t, J=6.7 Hz, 2H),1.84 (s, 3H), 1.78 (t, J=6.7 Hz, 2H), 1.31 (d, J=1.8 Hz, 6H); MS: (ES)m/z calculated for C₂₈H₂₇ClNO₄[M+H]⁺ 476.2, found 476.2.

Example 9: Synthesis of(S)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-1-(4-((5-oxo-4,5-dihydro-1H-tetrazol-1-yl)methyl)phenyl)indolin-2-one

Step a: To a solution of(S)-2-(4-(5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-2-oxoindolin-1-yl)phenyl)aceticacid (190.4 mg, 0.40 mmol) in dichloromethane (1.6 mL) at 0° C. undernitrogen, a solution of oxalyl chloride (52 μL, 0.60 mmol) indichloromethane (0.3 mL) was added dropwise. The reaction mixture wasstirred at 0° C. for 5 min and then warmed to room temperature andstirred for 4 hours. All solvents were removed under vacuum anddichloromethane (2 mL) was added to the residue. The mixture wasconcentrated under vacuum and this process was repeated another time togive the acid chloride product which was used in the next step directly.

Step b: To the acid chloride prepared in the previous step was addedazidotrimethylsilane (0.32 mL, 2.4 mmol) at room temperature (gasevolution!). The mixture was heated to 100° C. under nitrogen andstirred for 37 h. The reaction mixture was cooled to room temperatureand concentrated under vacuum. Water and dichloromethane were added tothe residue. The organic layer was separated, dried, and concentrated.The crude product was purified by silica gel chromatography (50% ethylacetate/hexane) to give the desired product. ¹H NMR (400 MHz,chloroform-d) δ 7.58-7.51 (m, 2H), 7.47-7.41 (m, 2H), 7.22-7.16 (m, 2H),7.03 (d, J=2.4 Hz, 1H), 6.96 (dd, J=8.6, 2.5 Hz, 1H), 6.85-6.80 (m, 1H),6.70 (d, J=8.6 Hz, 1H), 5.16 (s, 2H), 2.73 (t, J=6.7 Hz, 2H), 1.83 (s,3H), 1.77 (t, J=6.7 Hz, 2H), 1.30 (d, J=2.0 Hz, 6H). MS: (ES) m/zcalculated for C₂₈H₂₆ClN₅O₃ [M H]⁺516.2, found 516.5.

Example 10: Synthesis of(S)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-1-(4-((4-methyl-5-oxo-4,5-dihydro-1H-tetrazol-1-yl)methyl)phenyl)indolin-2-one

To a mixture of(S)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-1-(4-((5-oxo-4,5-dihydro-1H-tetrazol-1-yl)methyl)phenyl)indolin-2-one(204 mg, 0.395 mmol) and iodomethane (49 μL, 0.791 mmol) in DMF (1.0 mL)at 0° C. was added K₂CO₃ (138 mg, 1.0 mmol). The reaction mixture wasstirred at 0° C. for 20 min and then room temperature for 14 h. Themixture was poured into water (10 mL) and dichloromethane (3 mL). Theorganic layer was separated and the aqueous layer was extracted withdichloromethane. The combined organic layers were dried andconcentrated. The crude product was purified by silica gelchromatography (50% ethyl acetate/hexane) to give the desired product.¹H NMR (400 MHz, chloroform-d) δ 7.58-7.52 (m, 2H), 7.44-7.39 (m, 2H),7.21-7.16 (m, 2H), 7.03 (d, J=2.4 Hz, 1H), 6.98-6.93 (m, 1H), 6.85-6.80(m, 1H), 6.70 (d, J=8.6 Hz, 1H), 5.13 (s, 2H), 3.61 (s, 3H), 2.73 (t,J=6.7 Hz, 2H), 1.82 (s, 3H), 1.76 (t, J=6.7 Hz, 2H), 1.30 (d, J=1.8 Hz,6H). MS: (ES) m/z calculated for C₂₉H₂₈ClN₅O₃ [M+H]⁺ 530.2, found 530.5.

Compounds prepared by methods analogous to the methods described above,and evaluated using the serum chemotaxis assay below are provided in thefollowing table. A2 was calculated as described and activity ispresented in Table 1 as:

+, 20000 nM≥A₂≥500 nM; ++, 500 nM>A₂≥100 nM; +++, 100 nM>A₂.

TABLE 1 MS: (ES) Chemotaxis ¹H NMR m/z A₂

¹H NMR (400 MHz, Chloroform-d) δ 8.28-8.19 (m, 2H), 7.59 (dt, J = 8.4,0.5 Hz, 2H), 7.34 (d, J = 9.1 Hz, 2H), 7.31-7.27 (m, 2H), 7.17 (d, J =7.6 Hz, 1H), 7.03-6.94 (m, 1H), 6.85-6.78 (m, 1H), 2.36 (s, 3H), 1.89(s, 3H), 1.29 (s, 9H). 414.3 [M + H]⁺ ++

¹H NMR (400 MHz, Methanol-d₄) δ 8.28-8.17 (m, 2H), 7.66-7.56 (m, 2H),7.41 (d, J = 8.5 Hz, 2H), 7.31 (dd, J = 8.4, 2.2 Hz, 1H), 7.27 (s, 2H),7.25 (d, J = 0.6 Hz, 1H), 6.96 (d, J = 8.4 Hz, 1H), 1.88 (s, 3H), 1.30(s, 9H). 434.1 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.21-8.12 (m, 1H), 7.72 (dtd, J = 9.3,7.7, 1.6 Hz, 1H), 7.54 (td, J = 7.6, 1.1 Hz, 1H), 7.44-7.35 (m, 1H),7.36-7.32 (m, 2H), 7.32-7.27 (m, 2H), 7.24-7.14 (m, 2H), 6.53 (dd, J =17.9, 8.7 Hz, 1H), 1.76 (s, 3H), 1.28 (d, J = 12.5 Hz, 9H). 434.1 [M +H]⁺ ++

¹H NMR (400 MHz, Chloroform-d) δ 8.19-8.07 (m, 2H), 7.78-7.58 (m, 2H),7.21-7.16 (m, 1H), 7.13 (dd, J = 8.0, 1.9 Hz, 1H), 7.05 (d, J = 2.4 Hz,1H), 7.00 (dd, J = 8.6, 2.5 Hz, 1H), 6.88 (d, J = 1.8 Hz, 1H), 6.72 (d,J = 8.6 Hz, 1H), 2.74 (t, J = 6.7 Hz, 2H), 1.85 (s, 3H), 1.78 (t, J =6.7 Hz, 2H), 1.33-1.30 (m, 6H). 461.9 [M + H]⁺ ++

¹H NMR (400 MHz, Chloroform-d) δ 8.14 (dt, J = 4.1, 1.9 Hz, 2H),7.71-7.60 (m, 2H), 7.30-7.26 (m, 1H), 7.25 (q, J = 1.8 Hz, 2H),7.06-6.96 (m, 2H), 6.86 (d, J = 8.2 Hz, 1H), 1.89 (s, 3H), 1.36 (d, J =1.0 Hz, 9H). 474.1 [M + Na]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.20-8.06 (m, 2H), 7.71 (d, J = 7.9 Hz,1H), 7.66-7.57 (m, 1H), 7.35 (dd, J = 8.6, 1.2 Hz, 2H), 7.32-7.28 (m,2H), 7.11-7.03 (m, 2H), 6.83 (d, J = 8.0 Hz, 1H), 2.36 (s, 4H), 1.90 (d,J = 1.2 Hz, 3H), 1.30 (d, J = 1.2 Hz, 9H). 414.2 [M + H]⁺ ++

¹H NMR (400 MHz, Chloroform-d) δ 8.22 (d, J = 8.5 Hz, 2H), 7.60 (d, J =8.6 Hz, 2H), 7.35 (d, J = 8.6 Hz, 2H), 7.31-7.27 (m, 2H), 7.08 (d, J =10.0 Hz, 2H), 6.92 (d, J = 7.9 Hz, 1H), 2.36 (s, 3H), 1.89 (s, 3H), 1.29(d, J = 0.4 Hz, 9H). 414.2 [M + H]⁺ ++

¹H NMR (400 MHz, Chloroform-d) δ 8.33-8.24 (m, 2H), 7.65-7.60 (m, 2H),7.58 (d, J = 8.4 Hz, 1H), 7.35-7.29 (m, 1H), 7.25-7.19 (m, 1H), 7.10(dd, J = 2.3, 0.8 Hz, 1H), 6.98-6.88 (m, 2H), 1.90 (s, 3H), 1.84 (s,3H), 1.31 (d, J = 0.4 Hz, 9H). 448.1 [M + H]⁺ ++

¹H NMR (400 MHz, Chloroform-d) δ 8.20-8.14 (m, 2H), 7.77-7.71 (m, 1H),7.71-7.65 (m, 1H), 7.58 (d, J = 8.3 Hz, 1H), 7.35-7.31 (m, 1H), 7.20(dd, J = 8.4, 2.2 Hz, 1H), 7.13-7.09 (m, 1H), 6.94 (d, J = 2.1 Hz, 1H),6.84 (d, J = 8.4 Hz, 1H), 1.90 (s, 3H), 1.86 (s, 3H), 1.31 (s, 9H).448.1 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.18-8.09 (m, 2H), 7.71-7.66 (m, 1H),7.63 (dd, J = 8.2, 7.4 Hz, 1H), 7.27-7.24 (m, 2H), 7.23 (dq, J = 1.5,0.7 Hz, 2H), 6.88-6.80 (m, 3H), 4.52 (p, J = 6.1 Hz, 1H), 1.88 (s, 3H),1.32 (d, J = 6.0 Hz, 6H). 436.1 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.15 (dd, J = 9.0, 1.7 Hz, 1H), 8.02(d, J = 22.0 Hz, 1H), 7.62- 7.50 (m, 2H), 7.37 (d, J = 8.1 Hz, 3H), 7.24(dd, J = 1.6, 0.8 Hz, 2H), 7.14 (d, J = 2.0 Hz, 1H), 1.90 (s, 3H), 1.30(d, J = 0.5 Hz, 9H). 468.2 [M + H]⁺ +

¹H NMR (400 MHz, Chloroform-d) δ 8.21-8.12 (m, 1H), 7.72 (dtd, J = 9.3,7.7, 1.6 Hz, 1H), 7.54 (td, J = 7.6, 1.1 Hz, 1H), 7.44-7.36 (m, 1H),7.36-7.32 (m, 2H), 7.32- 7.27 (m, 2H), 7.24-7.14 (m, 2H), 6.53 (dd, J =17.9, 8.7 Hz, 1H), 1.76 (s, 3H), 1.28 (d, J = 12.5 Hz, 9H). 434.2 [M +H]⁺ ++

¹H NMR (400 MHz, Chloroform-d) δ 8.17 (s, 1H), 8.11 (d, J = 7.7 Hz, 1H),7.74-7.67 (m, 1H), 7.62 (t, J = 7.8 Hz, 1H), 7.25-7.23 (m, 1H),7.09-7.05 (m, 3H), 7.01 (dd, J = 13.9, 2.1 Hz, 1H), 6.83 (d, J = 8.4 Hz,1H), 2.37 (d, J = 0.9 Hz, 3H), 1.87 (d, J = 0.9 Hz, 3H), 1.35 (t, J =1.0 Hz, 9H). 432.2 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.20-8.08 (m, 2H), 7.68-7.63 (m, 2H),7.26-7.23 (m, 1H), 7.19 (d, J = 7.9 Hz, 1H), 7.16-7.11 (m, 1H),7.07-6.95 (m, 2H), 6.88 (d, J = 1.8 Hz, 1H), 1.87 (d, J = 0.7 Hz, 3H),1.35 (d, J = 0.9 Hz, 9H). 452.2 [M + H]⁺ +

¹H NMR (400 MHz, Chloroform-d) δ 8.29-8.22 (m, 2H), 7.61-7.54 (m, 2H),7.25-7.21 (m, 4H), 7.00- 6.93 (m, 1H), 6.87-6.80 (m, 2H), 4.61-4.38 (m,1H), 1.88 (s, 3H), 1.32 (dd, J = 6.1, 1.2 Hz, 6H). 436.2 [M + H]⁺ +++

452.2 [M + H]⁺ ++

452.1 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.18-8.12 (m, 2H), 7.72-7.68 (m, 1H),7.64 (t, J = 7.8 Hz, 1H), 7.25-7.21 (m, 1H), 7.16 (d, J = 7.6 Hz, 1H),7.10-7.05 (m, 1H), 7.04-6.96 (m, 2H), 6.71 (t, J = 1.2 Hz, 1H), 2.36 (d,J = 0.9 Hz, 3H), 1.87 (s, 3H), 1.35 (t, J = 0.7 Hz, 9H). 432.3 [M + H]⁺++

¹H NMR (400 MHz, Chloroform-d) δ 8.17 (t, J = 1.9 Hz, 1H), 8.12 (dt, J =7.5, 1.4 Hz, 1H), 7.71-7.67 (m, 1H), 7.63 (t, J = 7.8 Hz, 1H), 7.35-7.32(m, 2H), 7.32-7.28 (m, 2H), 7.17 (d, J = 7.6 Hz, 1H), 6.99-6.92 (m, 1H),6.72-6.69 (m, 1H), 2.35 (d, J = 0.7 Hz, 3H), 1.89 (s, 3H), 1.29 (d, J =0.5 Hz, 9H). 414.2 [M + H]⁺ ++

452.1 [M + H]⁺ ++

¹H NMR (400 MHz, DMSO-d₆) δ 8.86 (dd, J = 5.3, 0.8 Hz, 1H), 8.36-8.08(m, 1H), 7.84 (dd, J = 5.3, 2.1 Hz, 1H), 7.47 (d, J = 2.2 Hz, 1H), 7.38(d, J = 2.6 Hz, 1H), 7.36 (d, J = 2.4 Hz, 2H), 7.24 (d, J = 8.5 Hz, 2H),7.16 (d, J = 8.5 Hz, 1H), 1.84 (s, 3H), 1.23 (s, 9H). 435.0 [M + H]⁺ ++

¹H NMR (400 MHz, DMSO-d₆) δ 9.11 (d, J = 1.9 Hz, 1H), 8.97 (d, J = 2.4Hz, 1H), 8.40 (ddd, J = 2.5, 1.9, 0.6 Hz, 1H), 7.45 (d, J = 2.2 Hz, 1H),7.37 (d, J = 8.5 Hz, 2H), 7.32 (ddd, J = 8.5, 2.2, 0.6 Hz, 1H),7.30-7.26 (m, 2H), 6.95 (d, J = 8.5 Hz, 1H), 1.84 (s, 3H), 1.24 (d, J =0.6 Hz, 9H). 435.0 [M + H]⁺ ++

¹H NMR (400 MHz, DMSO-d₆) δ 8.81-8.77 (m, 1H), 8.20 (dd, J = 1.5, 0.8Hz, 1H), 7.87-7.78 (m, 1H), 7.71 (d, J = 8.6 Hz, 1H), 7.45 (d, J = 2.3Hz, 1H), 7.41-7.34 (m, 3H), 7.27-7.19 (m, 2H), 1.84 (s, 3H), 1.23 (s,9H). 435.0 [M + H]⁺ +++

¹H NMR (400 MHz, DMSO-d₆) δ 8.19-8.13 (m, 1H), 8.10-8.00 (m, 2H), 7.97(dt, J = 8.6, 0.5 Hz, 1H), 7.48-7.44 (m, 1H), 7.41 (ddd, J = 8.6, 2.3,0.6 Hz, 1H), 7.39-7.33 (m, 2H), 7.24-7.19 (m, 2H), 1.84 (s, 3H), 1.23(d, J = 0.6 Hz, 9H). 456.9 [M + Na]⁺ ++

¹H NMR (400 MHz, Chloroform-d) δ 8.14 (d, J = 5.4 Hz, 2H), 7.66 (d, J =7.1 Hz, 2H), 7.35-7.31 (m, 2H), 7.31 (d, J = 1.8 Hz, 2H), 7.29- 7.27 (m,1H), 7.23-7.21 (m, 1H), 6.86 (d, J = 8.4 Hz, 1H), 1.89 (s, 3H). 434.1[M + Na]⁺ +

¹H NMR (400 MHz, Chloroform-d) δ 7.84 (d, J = 64.1 Hz, 2H), 7.24- 7.13(m, 2H), 7.08 (d, J = 1.8 Hz, 1H), 7.03 (d, J = 7.8 Hz, 1H), 6.99- 6.87(m, 2H), 6.78 (t, J = 8.6 Hz, 1H), 6.63 (s, 1H), 3.74 (s, 3H), 1.63 (s,3H). 426.2 [M + H]⁺ +

¹H NMR (400 MHz, Chloroform-d) δ 8.17-8.11 (m, 2H), 7.69 (dt, J = 7.9,1.7 Hz, 1H), 7.64 (t, J = 8.0 Hz, 1H), 7.52-7.41 (m, 2H), 7.37-7.28 (m,2H), 7.25-7.22 (m, 1H), 6.86 (dd, J = 8.1, 0.8 Hz, 1H), 1.92 (s, 3H),1.57 (s, 6H). 457.9 [M + Na]⁺ ++

¹H NMR (400 MHz, DMSO-d₆) δ 8.03 (t, J = 8.3 Hz, 1H), 7.57 (dd, J =11.5, 1.9 Hz, 1H), 7.45 (td, J = 4.1, 1.9 Hz, 2H), 7.39-7.29 (m, 3H),7.28-7.22 (m, 2H), 7.02 (d, J = 8.5 Hz, 1H), 1.82 (s, 3H), 1.24 (d, J =0.6 Hz, 9H). 452.1 [M + H]⁺ ++

¹H NMR (400 MHz, Chloroform-d) δ 8.13 (t, J = 7.1 Hz, 1H), 7.76- 7.58(m, 1H), 7.47-7.32 (m, 3H), 7.32-7.26 (m, 2H), 7.26-7.19 (m, 2H), 6.63(dd, J = 8.6, 5.5 Hz, 1H), 1.92 (s, 3H), 1.30 (dd, J = 5.2, 1.0 Hz, 9H).452.1 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.16-8.12 (m, 2H), 7.72-7.61 (m, 2H),7.39-7.36 (m, 2H), 7.36- 7.32 (m, 2H), 7.24 (t, J = 1.1 Hz, 2H), 6.86(dd, J = 8.1, 0.9 Hz, 1H), 1.92 (d, J = 1.0 Hz, 3H), 1.67 (d, J = 21.9Hz, 6H). 459.9 [M + Na]⁺ ++

¹H NMR (400 MHz, Chloroform-d) δ 8.14 (d, J = 4.1 Hz, 2H), 7.74- 7.62(m, 2H), 7.57 (d, J = 8.5 Hz, 2H), 7.39 (d, J = 8.2 Hz, 2H), 7.26- 7.23(m, 2H), 6.87 (d, J = 8.1 Hz, 1H), 1.93 (s, 3H), 1.76 (s, 3H). 511.8[M + Na]⁺ ++

¹H NMR (400 MHz, Methanol-d₄) δ 8.17-8.11 (m, 1H), 8.10-8.05 (m, 1H),7.72 (d, J = 1.3 Hz, 1H), 7.71-7.70 (m, 1H), 7.42 (d, J = 2.0 Hz, 1H),7.41 (d, J = 2.0 Hz, 1H), 7.27 (d, J = 8.5 Hz, 2H), 7.22 (q, J = 1.0 Hz,2H), 7.00-6.94 (m, 1H), 1.91 (s, 3H), 1.31 (s, 9H). 484.2 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.14 (dt, J = 4.0, 1.8 Hz, 2H), 7.68(d, J = 2.1 Hz, 1H), 7.67- 7.62 (m, 1H), 7.44 (d, J = 8.1 Hz, 2H),7.35-7.31 (m, 2H), 7.25 (dt, J = 1.0, 0.5 Hz, 2H), 6.90-6.82 (m, 1H),1.91 (d, J = 0.8 Hz, 3H), 1.44-1.25 (m, 2H), 1.00 (s, 2H). 486.1 [M +H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.17-8.11 (m, 2H), 7.70-7.61 (m, 2H),7.37-7.32 (m, 2H), 7.29 (d, J = 2.2 Hz, 1H), 7.28-7.27 (m, 2H), 7.22 (d,J = 2.1 Hz, 1H), 6.86 (d, J = 8.4 Hz, 1H), 1.90 (s, 3H), 1.79-1.60 (m,2H), 1.44-1.27 (m, 2H). 443.1 [M + H]⁺ ++

¹H NMR (400 MHz, Chloroform-d) δ 8.17-8.09 (m, 2H), 7.68 (d, J = 8.2 Hz,1H), 7.66-7.60 (m, 1H), 7.24 (dtd, J = 2.6, 1.1, 0.6 Hz, 3H), 7.24-7.20(m, 3H), 6.85 (d, J = 9.0 Hz, 1H), 1.89 (s, 3H), 1.39 (s, 3H), 0.84 (s,2H), 0.78-0.64 (m, 2H). 432.2 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.24-8.09 (m, 2H), 7.71 (ddd, J = 8.0,2.1, 1.4 Hz, 1H), 7.65 (t, J = 8.0 Hz, 1H), 7.26 (dd, J = 2.6, 0.4 Hz,1H), 7.25-7.21 (m, 1H), 7.09- 7.06 (m, 1H), 7.05 (dt, J = 8.6, 1.6 Hz,1H), 6.84 (dt, J = 8.2, 0.5 Hz, 1H), 6.78 (d, J = 8.6 Hz, 1H), 4.49 (p,J = 6.0 Hz, 1H), 2.18 (s, 3H), 1.88 (s, 3H), 1.32 (dd, J = 6.0, 0.5 Hz,6H). 450.2 [M + H]⁺ ++

¹H NMR (400 MHz, Methanol-d₄) δ 7.60-7.56 (m, 2H), 7.56-7.53 (m, 3H),7.40-7.35 (m, 2H), 7.31- 7.29 (m, 1H), 7.28 (q, J = 1.9 Hz, 2H), 6.84(d, J = 9.1 Hz, 1H), 1.90 (s, 3H), 1.57 (s, 3H), 1.56 (s, 6H). 484.2[M + H]⁺ +

¹H NMR (400 MHz, Chloroform-d) δ 8.22-8.11 (m, 2H), 7.73-7.68 (m, 1H),7.65 (t, J = 7.7 Hz, 1H), 7.14 (d, J = 9.1 Hz, 2H), 7.05 (d, J = 2.4 Hz,1H), 6.99 (dd, J = 8.5, 2.6 Hz, 1H), 6.94-6.87 (m, 1H), 6.73 (d, J = 8.6Hz, 1H), 2.75 (t, J = 6.7 Hz, 2H), 1.88 (s, 3H), 1.78 (t, J = 6.7 Hz,2H), 1.32 (s, 6H). 512.2 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.21-8.16 (m, 1H), 8.15-8.10 (m, 1H),7.69-7.65 (m, 2H), 7.62- 7.54 (m, 1H), 7.53 (d, J = 1.6 Hz, 1H), 7.02(d, J = 2.5 Hz, 1H), 6.99- 6.94 (m, 2H), 6.74 (d, J = 8.6 Hz, 1H), 2.76(t, J = 6.8 Hz, 2H), 1.89 (s, 3H), 1.82-1.75 (m, 2H), 1.33 (d, J = 1.1Hz, 6H). 453.2 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.16 (dt, J = 10.3, 1.5 Hz, 1H),8.14-8.06 (m, 1H), 7.76-7.68 (m, 1H), 7.68-7.57 (m, 1H), 7.42- 7.33 (m,1H), 7.07 (dd, J = 14.5, 2.5 Hz, 1H), 7.02-6.93 (m, 2H), 6.81 (dd, J =8.3, 7.4 Hz, 1H), 6.75- 6.65 (m, 1H), 2.79-2.67 (m, 2H), 1.86 (s, 1H),1.85 (s, 2H), 1.78 (td, J = 6.7, 2.8 Hz, 2H), 1.32 (t, J = 1.7 Hz, 6H),0.99-0.92 (m, 1H), 0.69-0.56 (m, 1H). 468.2 [M + H]⁺ +++

¹H NMR (400 MHz, Methanol-d₄) δ 8.10 (ddd, J = 5.0, 3.5, 1.6 Hz, 1H),8.06 (d, J = 1.8 Hz, 1H), 7.73- 7.65 (m, 2H), 7.17-7.12 (m, 1H), 7.08(dd, J = 17.7, 2.0 Hz, 2H), 6.98 (dd, J = 8.6, 2.5 Hz, 1H), 6.81 (d, J =8.0 Hz, 1H), 6.66 (d, J = 8.6 Hz, 1H), 2.76 (t, J = 6.8 Hz, 2H), 2.64(q, J = 7.6 Hz, 2H), 1.83 (s, 3H), 1.79 (t, J = 6.8 Hz, 2H), 1.32-1.27(m, 6H), 1.21 (t, J = 7.6 Hz, 3H). 456.3 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.18-8.16 (m, 1H), 8.14 (dt, J = 7.8,1.4 Hz, 1H), 7.71 (ddd, J = 7.9, 2.1, 1.2 Hz, 1H), 7.67-7.61 (m, 1H),7.25-7.20 (m, 2H), 6.89- 6.82 (m, 3H), 2.73 (t, J = 6.8 Hz, 2H), 2.13(t, J = 0.6 Hz, 3H), 1.85 (s, 3H), 1.76 (t, J = 6.7 Hz, 2H), 1.31 (s,6H). 479.5 [M + H]⁺ +++

¹H NMR (400 MHz, Methanol-d₄) δ 8.12 (ddd, J = 4.9, 4.1, 1.6 Hz, 1H),8.05 (q, J = 1.2 Hz, 1H), 7.75- 7.58 (m, 2H), 7.37-7.20 (m, 2H),6.95-6.86 (m, 2H), 6.85- 6.79 (m, 1H), 2.79 (t, J = 6.8 Hz, 2H), 1.84(s, 3H), 1.82 (d, J = 6.8 Hz, 2H), 1.33 (s, 6H). 502.1 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.19-8.12 (m, 2H), 7.70 (ddt, J = 8.6,1.7, 0.9 Hz, 1H), 7.67-7.60 (m, 1H), 7.25-7.21 (m, 2H), 7.13 (t, J = 1.6Hz, 1H), 7.04 (dd, J = 8.5, 2.1 Hz, 1H), 6.85 (dd, J = 9.1, 1.1 Hz, 1H),6.73-6.65 (m, 1H), 3.00 (s, 2H), 1.87 (s, 3H), 1.69- 1.24 (m, 6H). 448.5[M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.07-7.92 (m, 2H), 7.57 (dt, J = 4.9,1.2 Hz, 2H), 7.43-7.29 (m, 1H), 7.25-7.14 (m, 2H), 7.11- 6.97 (m, 2H),6.87-6.71 (m, 1H), 2.52 (d, J = 3.9 Hz, 3H), 1.88 (d, J = 3.1 Hz, 3H),1.38 (dd, J = 1.3, 0.5 Hz, 9H). 448.3 [M + Na]⁺ ++

¹H NMR (400 MHz, Chloroform-d) δ 8.21-8.16 (m, 1H), 8.15 (tt, J = 1.8,0.8 Hz, 1H), 7.68-7.63 (m, 2H), 7.23 (q, J = 1.3 Hz, 1H), 7.03 (d, J =2.4 Hz, 1H), 7.01-6.96 (m, 2H), 6.74 (d, J = 8.5 Hz, 1H), 2.75 (t, J =6.6 Hz, 2H), 1.88 (s, 3H), 1.79 (t, J = 6.8 Hz, 2H), 1.32 (s, 6H). 546.3[M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.20-8.14 (m, 1H), 8.13 (q, J = 1.5 Hz,1H), 7.67 (d, J = 1.7 Hz, 1H), 7.66 (d, J = 1.6 Hz, 1H), 7.31 (s, 1H),7.05-6.98 (m, 2H), 6.97 (s, 1H), 6.74 (d, J = 8.6 Hz, 1H), 2.76 (t, J =6.8 Hz, 2H), 1.86 (s, 3H), 1.78 (d, J = 6.8 Hz, 2H), 1.32 (d, J = 1.5Hz, 6H). 518.2 [M + Na]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.18-8.10 (m, 2H), 7.72-7.59 (m, 2H),7.40-7.35 (m, 2H), 7.29 (d, J = 2.1 Hz, 1H), 7.28-7.26 (m, 1H), 7.24 (d,J = 7.7 Hz, 2H), 6.87- 6.81 (m, 1H), 1.91 (s, 3H), 1.30 (s, 9H). 434.2[M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.08 (dd, J = 6.2, 2.7 Hz, 1H), 7.65(ddd, J = 8.8, 4.0, 2.7 Hz, 1H), 7.44-7.29 (m, 3H), 7.28-7.20 (m, 4H),6.81 (d, J = 9.0 Hz, 1H), 1.90 (s, 3H), 1.30 (s, 9H). 452.2 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.13-8.06 (m, 1H), 7.51 (dd, J = 8.1,2.2 Hz, 1H), 7.42 (d, J = 8.2 Hz, 1H), 7.39-7.31 (m, 2H), 7.30- 7.17 (m,4H), 6.85-6.78 (m, 1H), 2.69 (s, 3H), 1.89 (s, 3H), 1.30 (s, 9H). 448.2[M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.22-8.09 (m, 2H), 7.69 (dddd, J = 8.0,1.8, 1.3, 0.4 Hz, 1H), 7.66- 7.58 (m, 1H), 7.26-7.17 (m, 2H), 7.06 (d, J= 2.3 Hz, 1H), 6.99 (ddd, J = 8.5, 2.5, 0.6 Hz, 1H), 6.88-6.81 (m, 1H),6.73 (d, J = 8.6 Hz, 1H), 2.76 (t, J = 6.7 Hz, 2H), 1.87 (s, 3H), 1.78(t, J = 6.7 Hz, 2H), 1.32 (d, J = 1.5 Hz, 6H). 462.5 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.10 (d, J = 2.3 Hz, 1H), 7.52 (dd, J =8.1, 2.3 Hz, 1H), 7.42 (dt, J = 8.0, 0.7 Hz, 1H), 7.22-7.19 (m, 2H),7.06-7.01 (m, 1H), 6.98 (ddd, J = 8.7, 2.5, 0.7 Hz, 1H), 6.83 (dd, J =8.9, 0.6 Hz, 1H), 6.73 (d, J = 8.5 Hz, 1H), 2.80-2.71 (m, 2H), 2.69 (s,3H), 1.85 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32 (d, J = 1.6 Hz, 6H).476.2 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.39-8.13 (m, 2H), 7.65-7.50 (m, 2H),7.30-7.25 (m, 2H), 7.05 (d, J = 2.3 Hz, 1H), 7.00-6.92 (m, 2H), 6.73 (d,J = 8.6 Hz, 1H), 2.75 (t, J = 6.8 Hz, 2H), 1.86 (s, 3H), 1.78 (t, J =6.7 Hz, 2H), 1.43-1.08 (m, 6H). 462.2 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.08 (dd, J = 6.3, 2.7 Hz, 1H), 7.66(ddd, J = 8.8, 4.0, 2.7 Hz, 1H), 7.32 (dd, J = 9.9, 8.8 Hz, 1H), 7.27-7.23 (m, 2H), 7.04 (d, J = 2.5 Hz, 1H), 6.97 (ddd, J = 8.7, 2.5, 0.6 Hz,1H), 6.87-6.78 (m, 1H), 6.73 (d, J = 8.6 Hz, 1H), 2.95-2.40 (m, 2H),1.86 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.47-1.17 (m, 6H). 480.2 [M +H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.08 (d, J = 2.5 Hz, 1H), 7.63 (d, J =8.6 Hz, 1H), 7.57 (ddd, J = 8.6, 2.5, 0.6 Hz, 1H), 7.27-7.22 (m, 2H),7.04 (d, J = 2.4 Hz, 1H), 6.96 (ddd, J = 8.7, 2.5, 0.7 Hz, 1H), 6.86(dt, J = 8.1, 0.6 Hz, 1H), 6.72 (d, J = 8.6 Hz, 1H), 2.75 (t, J = 6.8Hz, 2H), 1.85 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32-1.30 (m, 6H).496.5 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.17 (d, J = 8.4 Hz, 1H), 7.65 (d, J =2.0 Hz, 1H), 7.49 (dd, J = 8.5, 2.1 Hz, 1H), 7.29-7.19 (m, 2H), 7.03 (d,J = 2.4 Hz, 1H), 7.00- 6.93 (m, 2H), 6.73 (d, J = 8.6 Hz, 1H), 2.75 (t,J = 6.8 Hz, 2H), 1.86 (s, 3H), 1.79 (t, J = 6.7 Hz, 2H), 1.32 (s, 6H).496.5 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.21 (d, J = 2.7 Hz, 1H), 7.66 (dd, J =8.9, 2.7 Hz, 1H), 7.29-7.16 (m, 3 H), 7.01 (d, J = 2.4 Hz, 1H), 6.96(dd, J = 8.6, 2.5 Hz, 1H), 6.78 (dd, J = 8.1, 0.7 Hz, 1H), 6.73 (d, J =8.6 Hz, 1H), 4.13 (s, 3H), 2.75 (t, J = 6.7 Hz, 2H), 1.85 (s, 3H), 1.78(t, J = 6.7 Hz, 2H), 1.32 (s, 6H). 492.3 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.47 (t, J = 7.7 Hz, 1H), 7.41- 7.30(m, 3H), 7.22-7.16 (m, 2H), 7.05 (d, J = 2.4 Hz, 1H), 6.97 (dd, J = 8.6,2.4 Hz, 1H), 6.85 (d, J = 8.8 Hz, 1H), 6.71 (d, J = 8.6 Hz, 1H), 3.71(s, 2H), 2.74 (t, J = 6.8 Hz, 2H), 1.84 (s, 3H), 1.77 (t, J = 6.7 Hz,2H), 1.31 (d, J = 1.8 Hz, 6H). 476.6 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.12 (d, J = 2.1 Hz, 1H), 7.54 (dd, J =8.2, 2.3 Hz, 1H), 7.41 (d, J = 8.2 Hz, 1H), 7.12-7.02 (m, 3H), 7.00 (dd,J = 8.6, 2.4 Hz, 1H), 6.80 (d, J = 7.8 Hz, 1H), 6.71 (d, J = 8.6 Hz,1H), 2.75 (t, J = 6.8 Hz, 2H), 2.69 (s, 3H), 2.35 (s, 3H), 1.84 (s, 3H),1.77 (t, J = 6.7 Hz, 2H), 1.37-1.27 (m, 6H). 456.0 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.19 (t, J = 1.9 Hz, 1H), 8.13- 8.06(m, 1H), 7.76-7.69 (m, 1H), 7.61 (t, J = 7.9 Hz, 1H), 7.14-7.03 (m, 3H),7.03-6.97 (m, 1H), 6.86- 6.77 (m, 1H), 6.71 (d, J = 8.6 Hz, 1H), 2.75(t, J = 6.8 Hz, 2H), 2.36 (d, J = 0.9 Hz, 3H), 1.85 (s, 3H), 1.78 (t, J= 6.7 Hz, 2H), 1.31 (d, J = 1.8 Hz, 6H). 442.5 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.26-8.20 (m, 2H), 7.61 (d, J = 8.6 Hz,2H), 7.08 (dt, J = 6.3, 3.4 Hz, 3H), 6.98 (dd, J = 8.6, 2.5 Hz, 1H),6.92 (d, J = 8.5 Hz, 1H), 6.71 (d, J = 8.6 Hz, 1H), 2.75 (t, J = 6.8 Hz,2H), 2.36 (s, 3H), 1.85 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.31 (d, J =1.7 Hz, 6H). 442.5 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.17 (d, J = 2.7 Hz, 1H), 7.73 (dd, J =8.8, 2.7 Hz, 1H), 7.51 (dd, J = 8.8, 1.4 Hz, 1H), 7.29-7.23 (m, 2H),7.07-7.00 (m, 1H), 6.97 (dd, J = 8.6, 2.5 Hz, 1H), 6.90 (dd, J = 8.2,0.6 Hz, 1H), 6.73 (d, J = 8.6 Hz, 1H), 2.74 (d, J = 6.8 Hz, 2H), 1.86(s, 3H), 1.78 (s, 2H), 1.32 (d, J = 1.0 Hz, 6H). 546.6 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.49-7.41 (m, 2H), 7.41-7.34 (m, 2H),7.20 (dq, J = 4.3, 2.1 Hz, 2H), 7.04 (d, J = 2.3 Hz, 1H), 6.96 (ddd, J =8.6, 1.9, 1.2 Hz, 1H), 6.85 (d, J = 8.9 Hz, 1H), 6.71 (d, J = 8.5 Hz,1H), 3.71 (s, 2H), 2.74 (t, J = 6.7 Hz, 2H), 1.84 (s, 3H), 1.78 (t, J =6.7 Hz, 2H), 1.31 (d, J = 1.8 Hz, 6H). 476.2 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.18 (s, 1H), 7.44-7.35 (m, 2H), 7.28(dd, J = 8.4, 2.1 Hz, 1H), 7.25 (d, J = 2.1 Hz, 1H), 7.04-7.00 (m, 2H),6.95 (dd, J = 8.6, 2.4 Hz, 1H), 6.73 (d, J = 8.6 Hz, 1H), 2.75 (t, J =6.8 Hz, 2H), 1.86 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32 (d, J = 1.1Hz, 6H). 480.5 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.19 (t, J = 1.8 Hz, 1H), 8.10 (dt, J =7.8, 1.4 Hz, 1H), 7.72 (ddd, J = 7.9, 2.1, 1.2 Hz, 1H), 7.62 (t, J = 7.9Hz, 1H), 7.11-7.04 (m, 3H), 7.01 (dd, J = 8.6, 2.5 Hz, 1H), 6.83 (d, J =7.9 Hz, 1H), 6.71 (d, J = 8.6 Hz, 1H), 2.75 (t, J = 6.7 Hz, 2H), 2.36(s, 3H), 1.85 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.31 (d, J = 1.8 Hz,6H). 442.6 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.28-8.17 (m, 1H), 7.40-7.35 (m, 2H),7.27-7.23 (m, 2H), 7.06 (d, J = 2.4 Hz, 1H), 6.97 (dd, J = 8.6, 2.5 Hz,1H), 6.95-6.89 (m, 1H), 6.73 (d, J = 8.6 Hz, 1H), 2.76 (t, J = 6.8 Hz,2H), 2.71 (s, 3H), 1.86 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32 (d, J =1.6 Hz, 6H). 476.2 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.39-7.30 (m, 4H), 7.22-7.15 (m, 2H),7.05 (d, J = 2.4 Hz, 1H), 6.99-6.94 (m, 1H), 6.85-6.80 (m, 1H), 6.71 (d,J = 8.6 Hz, 1H), 3.02 (t, J = 7.7 Hz, 2H), 2.81-2.68 (m, 4H), 1.83 (s,3H), 1.78 (t, J = 6.7 Hz, 2H), 1.31 (d, J = 1.9 Hz, 6H). 490.2 [M + H]⁺+++

¹H NMR (400 MHz, CDCl₃) δ 8.05 (d, J = 2.3, Hz, 1 H), 7.56 (dd, J = 8.5,2.3 Hz, 1 H), 7.46 (d, J = 8.2 Hz, 1 H), 7.23-7.20 (m, 2 H), 7.05 (d, J= 2.4, Hz, 1 H), 6.98 (dd, J = 8.6, 2.3 Hz, 1 H) 6.85-6.83 (m, 1 H),6.73 (d, J = 8.6 Hz, 1 H), 3.11 (q, J = 7.4 Hz, 2 H), 3.07 (bs, 1 H),2.76 (t, J = 7.6 Hz, 2 H), 1.84 (s, 3 H), 1.79 (t, J = 6.6 Hz, 2 H),1.31-1.25 (m, 9 H). 490.2 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.30 (dd, J = 8.8, 0.6 Hz, 1H),7.31-7.19 (m, 4H), 7.07-7.02 (m, 1H), 7.00 (dt, J = 8.4, 0.5 Hz, 1H),6.95 (ddd, J = 8.7, 2.5, 0.7 Hz, 1H), 6.73 (d, J = 8.6 Hz, 1H), 4.09 (s,3H), 2.86-2.52 (m, 2H), 1.86 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H),1.34-1.29 (m, 6H). 492.2 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.49-7.39 (m, 1H), 7.25 (ddd, J = 7.5,5.3, 3.1 Hz, 3H), 7.20-7.18 (m, 1H), 7.05 (d, J = 2.4 Hz, 1H), 6.97 (dd,J = 8.6, 2.5 Hz, 1H), 6.85- 6.79 (m, 1H), 6.72 (d, J = 8.6 Hz, 1H), 3.00(t, J = 7.7 Hz, 2H), 2.73 (dt, J = 17.0, 7.2 Hz, 4H), 1.84 (s, 3H), 1.78(t, J = 6.7 Hz, 2H), 1.31 (d, J = 1.9 Hz, 6H). 490.2 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.06-7.99 (m, 2H), 7.51-7.35 (m, 2H),7.20-7.07 (m, 2H), 7.00- 6.82 (m, 2H), 6.71 (s, 1H), 6.66 (d, J = 8.9Hz, 1H), 5.11-4.75 (m, 2H), 2.74 (t, J = 6.7 Hz, 2H), 1.82 (s, 3H), 1.78(t, J = 6.7 Hz, 2H), 1.31 (s, 6H). 476.2 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.47 (t, J = 7.8 Hz, 1H), 7.42- 7.29(m, 3H), 7.22-7.16 (m, 2H), 7.05 (dd, J = 2.5, 1.2 Hz, 1H), 7.00- 6.94(m, 1H), 6.84 (dt, J = 8.0, 1.0 Hz, 1H), 6.72 (d, J = 8.6 Hz, 1H), 3.80(d, J = 7.4 Hz, 1H), 2.75 (t, J = 6.7 Hz, 2H), 1.84 (d, J = 1.0 Hz, 3H),1.78 (t, J = 6.7 Hz, 2H), 1.55 (d, J = 7.0 Hz, 3H), 1.31 (d, J = 1.8 Hz,6H). 490.6 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.05 (d, J = 8.2 Hz, 2H), 7.34 (d, J =8.1 Hz, 2H), 7.19-7.12 (m, 2H), 6.99 (d, J = 2.4 Hz, 1H), 6.91 (dd, J =8.6, 2.5 Hz, 1H), 6.72 (d, J = 8.5 Hz, 1H), 6.67-6.60 (m, 1H), 5.09-4.89(m, 2H), 2.74 (t, J = 6.8 Hz, 2H), 1.81 (s, 3H), 1.78 (t, J = 6.7 Hz,2H), 1.32 (s, 6H). 476.2 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.16 (dd, J = 7.8, 1.5 Hz, 1H),7.53-7.43 (m, 1H), 7.42-7.33 (m, 1H), 7.21-7.12 (m, 2H), 7.07- 6.99 (m,2H), 6.95 (dd, J = 8.6, 2.5 Hz, 1H), 6.72 (d, J = 8.5 Hz, 1H), 6.66 (dd,J = 8.3, 0.6 Hz, 1H), 5.44 (d, J = 3.8 Hz, 2H), 2.75 (t, J = 6.7 Hz,2H), 1.84 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32 (s, 6H). 476.2 [M +H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.53-7.42 (m, 3H), 7.34-7.28 (m, 1H),7.22-7.17 (m, 2H), 7.08- 7.03 (m, 1H), 6.97 (dd, J = 8.6, 2.5 Hz, 1H),6.82 (dd, J = 8.1, 0.8 Hz, 1H), 6.72 (d, J = 8.6 Hz, 1H), 2.75 (t, J =6.8 Hz, 2H), 1.85 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.63 (s, 6H), 1.32(d, J = 2.1 Hz, 6H). 504.2 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.17 (d, J = 2.4 Hz, 1H),7.45- 7.37 (m,1H), 7.20-7.10 (m, 2H), 7.03-6.96 (m, 2H), 6.90-6.83 (m, 1H), 6.76-6.63(m, 2H), 4.98- 4.82 (m, 2H), 4.06 (s, 3H), 2.75 (t, J = 6.7 Hz, 2H),1.81-1.74 (m, 5H), 1.32 (s, 6H). 528.2 [M + Na]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.08 (s, 1H), 8.02 (d, J = 7.7 Hz, 1H),7.60-7.37 (m, 2H), 7.11 (s, 1H), 7.06-6.96 (m, 2H), 6.92- 6.85 (m, 1H),6.77-6.65 (m, 1H), 6.41 (t, J = 8.6 Hz, 1H), 5.87 (m, J = 7.0 Hz, 1H),2.76 (d, J = 7.0 Hz, 2H), 1.87 (dd, J = 7.1, 4.0 Hz, 2H), 1.81-1.74 (m,6H), 1.32 (d, J = 2.3 Hz, 6H). 512.2 [M + Na]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.92 (d, J = 6.8 Hz, 1H), 7.44 (s, 1H),7.22-7.08 (m, 3H), 6.97 (s, 1H), 6.89 (d, J = 10.7 Hz, 1H), 6.68 (dd, J= 16.5, 8.4 Hz, 2H), 5.06- 4.79 (m, 2H), 2.73 (t, J = 6.7 Hz, 3H),1.81-1.68 (m, 5H), 1.31 (s, 6H). 516.3 [M + Na]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.93 (s, 1H), 7.33-7.27 (m, 1H), 7.22(d, J = 7.0 Hz, 1H), 7.18- 7.12 (m, 2H), 6.99 (d, J = 4.2 Hz, 1H),6.92-6.86 (m, 1H), 6.72- 6.64 (m, 2H), 5.03-4.78 (m, 2H), 2.74 (t, J =6.7 Hz, 2H), 2.60 (s, 3H), 1.80 (m, 5H), 1.31 (s, 6H). 512.3 [M + Na]⁺+++

¹H NMR (400 MHz, Chloroform-d) δ 7.92 (s, 1H), 7.25-7.13 (m, 3H), 7.00(d, J = 4.5 Hz, 1H), 6.97- 6.90 (m, 2H), 6.69 (dd, J = 17.2, 8.3 Hz,2H), 5.35 (d, J = 5.1 Hz, 2H), 2.74 (t, J = 6.8 Hz, 2H), 2.36 (s, 3H),1.81 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32 (s, 6H). 490.3 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.42 (t, J = 8.1 Hz, 1H), 7.22- 7.16(m, 2H), 7.08-7.02 (m, 2H), 7.01-6.92 (m, 3H), 6.86 (dd, J = 9.0, 0.6Hz, 1H), 6.71 (d, J = 8.6 Hz, 1H), 4.66 (s, 2H), 2.74 (t, J = 6.7 Hz,2H), 1.83 (s, 3H), 1.77 (t, J = 6.7 Hz, 2H), 1.31 (d, J = 1.8 Hz, 6H).492.5 [M + H]⁺ +++

¹H NMR (400 MHz, Methanol-d₄) δ 7.77 (dd, J = 9.5, 2.8 Hz, 1H),7.33-7.17 (m, 2H), 7.12-7.03 (m, 1H), 6.99 (d, J = 2.4 Hz, 1H), 6.94 (d,J = 8.4 Hz, 1H), 6.87- 6.80 (m, 2H), 6.67 (d, J = 8.6 Hz, 1H), 5.38 (s,2H), 2.76 (t, J = 6.6 Hz, 2H), 1.85-1.75 (m, 5H), 1.29 (d, J = 1.9 Hz,6H). 494.3 [M + H]⁺ +++

¹H NMR (400 MHz, Methanol-d₄) δ 7.58 (d, J = 2.6 Hz, 1H), 7.24- 7.19 (m,3H), 7.04-6.97 (m, 2H), 6.97-6.90 (m, 1H), 6.82 (d, J = 8.6 Hz, 1H),6.67 (d, J = 8.5 Hz, 1H), 5.33 (s, 2H), 3.80 (s, 3H), 2.76 (t, J = 6.8Hz, 2H), 1.88-1.67 (m, 5H), 1.29 (d, J = 2.0 Hz, 6H). 506.4 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.32 (d, J = 8.8 Hz, 2H), 7.23- 7.16(m, 2H), 7.06-7.01 (m, 3H), 6.97-6.88 (m, 1H), 6.80-6.74 (m, 1H), 6.72(d, J = 8.6 Hz, 1H), 4.69 (s, 2H), 2.74 (s, 2H), 1.84 (s, 3H), 1.78 (t,J = 6.7 Hz, 2H), 1.31 (d, J = 1.8 Hz, 6H). 492.5 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.37-7.28 (m, 4H), 7.20 (dqd, J = 4.2,2.2, 0.6 Hz, 2H), 7.05 (d, J = 2.4 Hz, 1H), 6.96 (ddd, J = 8.8, 2.5, 0.8Hz, 1H), 6.81 (dd, J = 8.9, 0.6 Hz, 1H), 6.72 (d, J = 8.5 Hz, 1H),2.79-2.65 (m, 4H), 2.42 (t, J = 7.3 Hz, 2H), 2.00 (p, J = 7.4 Hz, 2H),1.84 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32 (d, J = 2.0 Hz, 6H). 504.6[M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.23 (d, J = 2.7 Hz, 1H), 7.64 (ddd, J= 8.9, 2.8, 0.6 Hz, 1H), 7.25- 7.15 (m, 3H), 7.01 (d, J = 2.4 Hz, 1H),6.96 (ddd, J = 8.7, 2.5, 0.7 Hz, 1H), 6.78 (dt, J = 8.1, 0.7 Hz, 1H),6.73 (d, J = 8.5 Hz, 1H), 4.40 (q, J = 7.0 Hz, 2H), 2.75 (t, J = 6.7 Hz,2H), 1.85 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.61 (t, J = 7.0 Hz, 3H),1.32 (s, 6H). 506.4 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.25 (d, J = 2.7 Hz, 1H), 7.63 (dd, J =8.8, 2.8 Hz, 1H), 7.25-7.16 (m, 3H), 7.06-6.92 (m, 2H), 6.80 (d, J = 9.0Hz, 1H), 6.72 (d, J = 8.5 Hz, 1H), 4.93 (p, J = 6.1 Hz, 1H), 2.75 (t, J= 6.7 Hz, 2H), 1.84 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.53 (dd, J =6.0, 1.0 Hz, 6H), 1.32 (s, 6H). 520.4 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.19-8.05 (m, 1H), 7.39 (dd, J = 8.4,2.3 Hz, 1H), 7.23-7.16 (m, 2H), 7.03-6.96 (m, 2H), 6.92 (dd, J = 8.5,2.5 Hz, 1H), 6.72 (d, J = 8.6 Hz, 1H), 6.65 (d, J = 8.1 Hz, 1H),5.43-5.24 (m, 2H), 2.74 (t, J = 6.8 Hz, 2H), 2.40 (br s, 1H), 1.88- 1.64(m, 5H), 1.32 (s, 6H). 510.3 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.97 (d, J = 7.9 Hz, 1H), 7.24 (s, 1H),7.18-7.13 (m, 2H), 7.10 (s, 1H), 6.98 (s, 1H), 6.91 (d, J = 8.6 Hz, 1H),6.71 (d, J = 8.6 Hz, 1H), 6.63 (d, J = 8.8 Hz, 1H), 5.09- 4.65 (m, 2H),2.77-2.70 (m, 2H), 2.57 (d, J = 0.8 Hz, 3H), 1.87- 1.61 (m, 5H), 1.32(s, 6H). 490.4 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.05-8.00 (m, 1H), 7.19-7.10 (m, 3H),7.06-7.02 (m, 1H), 6.97- 6.92 (m, 1H), 6.82-6.79 (m, 1H), 6.74-6.64 (m,1H), 5.38 (d, J = 21.9 Hz, 2H), 2.79-2.74 (m, 2H), 2.22 (s, 3H), 1.83(s, 3H), 1.79- 1.74 (m, 2H), 1.32 (d, J = 3.2 Hz, 6H). 490.4 [M + H]⁺+++

¹H NMR (400 MHz, Chloroform-d) δ 7.26-7.25 (m, 1H), 7.25-7.21 (m, 1H),7.20 (d, J = 9.1 Hz, 1H), 7.18-7.13 (m, 1H), 6.99 (d, J = 7.6 Hz, 1H),6.91 (d, J = 8.4 Hz, 1H), 6.85 (d, J = 8.1 Hz, 2H), 6.68 (d, J = 8.1 Hz,1H), 5.11-4.71 (m, 2H), 2.71-2.66 (m, 2H), 2.44 (s, 3H), 1.77-1.73 (m,5H), 1.30 (s, 6H). 512.3 [M + Na]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.75 (t, J = 1.6 Hz, 1H), 7.65 (dd, J =2.5, 1.3 Hz, 1H), 7.26-7.19 (m, 3H), 7.05 (d, J = 2.5 Hz, 1H), 6.99 (dd,J = 8.6, 2.5 Hz, 1H), 6.90- 6.84 (m, 1H), 6.73 (d, J = 8.6 Hz, 1H), 3.89(s, 3H), 2.76 (t, J = 6.7 Hz, 2H), 1.86 (s, 3H), 1.78 (t, J = 6.7 Hz,2H), 1.32 (d, J = 1.6 Hz, 6H). 492.5 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.95 (dd, J = 1.8, 0.8 Hz, 2H), 7.50(dq, J = 2.0, 0.9 Hz, 1H), 7.22 (dt, J = 4.3, 2.4 Hz, 2H), 7.06 (d, J =2.4 Hz, 1H), 6.99 (dd, J = 8.6, 2.4 Hz, 1H), 6.83 (d, J = 9.0 Hz, 1H),6.73 (d, J = 8.6 Hz, 1H), 2.76 (t, J = 6.8 Hz, 2H), 2.48 (s, 3H), 1.86(s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32 (d, J = 1.6 Hz, 6H). 476.3 [M +H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.09 (d, J = 2.1 Hz, 1H), 7.94 (d, J =8.5 Hz, 1H), 7.79 (ddd, J = 8.5, 2.0, 0.8 Hz, 1H), 7.31-7.22 (m, 2H),7.03 (d, J = 2.4 Hz, 1H), 7.01- 6.90 (m, 2H), 6.73 (d, J = 8.6 Hz, 1H),2.75 (t, J = 6.8 Hz, 2H), 1.87 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32(s, 6H). 530.3 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.86 (d, J = 7.7 Hz, 1H), 7.20- 7.08(m, 4H), 7.03 (d, J = 2.4 Hz, 1H), 6.94 (dd, J = 8.6, 2.5 Hz, 1H), 6.72(d, J = 8.6 Hz, 1H), 6.54 (d, J = 8.3 Hz, 1H), 4.98 (d, J = 5.6 Hz, 2H),2.75 (t, J = 6.8 Hz, 2H), 2.62 (s, 3H), 1.83 (s, 3H), 1.78 (t, J = 6.8Hz, 2H), 1.32 (s, 6H). 490.3 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.63 (d, J = 7.2 Hz, 1H), 7.35- 7.28(m, 2H), 7.25-7.18 (m, 1H), 7.17 (dd, J = 2.0, 1.1 Hz, 1H), 6.84 (dd, J= 13.4, 5.0 Hz, 3H), 6.67 (d, J = 8.5 Hz, 1H), 5.11 (s, 2H), 2.74- 2.47(m, 2H), 2.18 (d, J = 1.0 Hz, 3H), 1.76 (t, J = 6.6 Hz, 2H), 1.72 (d, J= 1.1 Hz, 3H), 1.30 (s, 6H). . 490.4 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.48-7.41 (m, 3H), 7.30-7.23 (m, 2H),7.20-7.12 (m, 2H), 7.03 (dd, J = 15.9, 2.5 Hz, 1H), 6.94 (ddd, J = 12.7,8.5, 2.5 Hz, 1H), 6.70 (dd, J = 8.6, 7.0 Hz, 1H), 6.55 (dd, J = 26.1,8.3 Hz, 1H), 3.74- 3.30 (m, 2H), 2.72 (q, J = 6.7 Hz, 2H), 1.84 (s, 3H),1.77 (q, J = 6.8 Hz, 2H), 1.31 (d, J = 2.0 Hz, 6H) 476.3 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.23 (d, J = 2.7 Hz, 1H), 7.63 (dd, J =8.8, 2.7 Hz, 1H), 7.23-7.17 (m, 3H), 7.02 (d, J = 2.4 Hz, 1H), 6.97 (dd,J = 8.6, 2.4 Hz, 1H), 6.81- 6.75 (m, 1H), 6.73 (d, J = 8.5 Hz, 1H), 4.08(d, J = 6.4 Hz, 2H), 2.75 (t, J = 6.7 Hz, 2H), 2.27 (dt, J = 13.3, 6.7Hz, 1H), 1.84 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32 (s, 6H), 1.28 (s,2H), 1.12 (d, J = 6.7 Hz, 6H). 534.4 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.09 (d, J = 8.4 Hz, 1H), 7.34 (dd, J =8.4, 2.1 Hz, 1H), 7.24-7.16 (m, 2H), 7.04-6.89 (m, 3H), 6.75 (d, J = 8.5Hz, 1H), 6.65 (d, J = 9.1 Hz, 1H), 5.45 (d, J = 17.9 Hz, 1H), 5.32 (d, J= 17.9 Hz, 1H), 2.75 (t, J = 6.7 Hz, 2H), 1.84 (s, 3H), 1.78 (t, J = 6.7Hz, 2H), 1.32 (d, J = 1.4 Hz, 6H). 532.3 [M + Na]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.08 (dt, J = 12.0, 1.6 Hz, 2H), 7.71(t, J = 1.9 Hz, 1H), 7.28-7.26 (m, 1H), 7.24 (t, J = 1.6 Hz, 1H), 7.04(d, J = 2.4 Hz, 1H), 6.97 (dd, J = 8.6, 2.5 Hz, 1H), 6.88 (d, J = 8.2Hz, 1H), 6.73 (d, J = 8.5 Hz, 1H), 2.76 (t, J = 6.7 Hz, 2H), 1.86 (s,3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32 (d, J = 1.2 Hz, 6H). 496.3 [M + H]⁺+++

¹H NMR (400 MHz, Chloroform-d) δ 7.37 (s, 1H), 7.28-7.27 (m, 1H),7.24-7.21 (m, 1H), 7.16 (d, J = 2.1 Hz, 1H), 7.05 (d, J = 7.7 Hz, 1H),6.92 (d, J = 2.4 Hz, 1H), 6.86 (d, J = 8.4 Hz, 2H), 6.69 (d, J = 8.6 Hz,1H), 4.96 (d, J = 2.0 Hz, 2H), 2.88-2.53 (m, 2H), 1.79-1.66 (m, 5H),1.31 (s, 6H). 532.3 [M + Na]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.42-7.31 (m, 2H), 7.31-7.27 (m, 1H),7.18 (d, J = 8.5 Hz, 1H), 7.10 (d, J = 2.1 Hz, 1H), 6.94 (d, J = 8.1 Hz,1H), 6.74 (d, J = 2.4 Hz, 1H), 6.69 (dd, J = 8.6, 2.5 Hz, 1H), 6.60 (d,J = 8.6 Hz, 1H), 4.96 (d, J = 3.4 Hz, 2H), 3.58 (s, 3H), 2.64 (q, J =6.5 Hz, 2H), 1.73 (t, J = 6.7 Hz, 2H), 1.68 (s, 3H), 1.28 (s, 6H). 506.3[M + H]⁺ ++

¹H NMR (400 MHz, Chloroform-d) δ 7.33 (t, J = 8.2 Hz, 2H), 7.21- 7.13(m, 2H), 7.03-6.83 (m, 2H), 6.76-6.65 (m, 3H), 5.22 (s, 2H), 4.00 (s,3H), 2.73 (t, J = 6.8 Hz, 2H), 1.80-1.73 (m, 5H), 1.31 (s, 6H). 506.3[M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 7.99 (t, J = 1.6 Hz, 1H), 7.81 (ddd, J= 8.4, 2.5, 1.4 Hz, 1H), 7.48 (ddd, J = 8.8, 2.4, 1.8 Hz, 1H), 7.28-7.23(m, 2H), 7.03 (dd, J = 2.4, 1.1 Hz, 1H), 6.97 (dd, J = 8.6, 2.5 Hz, 1H),6.92 (dd, J = 8.3, 0.5 Hz, 1H), 6.73 (d, J = 8.5 Hz, 1H), 2.76 (t, J =6.8 Hz, 2H), 1.86 (s, 3H), 1.79 (t, J = 6.7 Hz, 2H), 1.35-1.16 (m, 6H).480.3 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.13 (d, J = 8.7 Hz, 1H), 7.22- 7.15(m, 2H), 7.05 (s, 1H), 6.97 (d, J = 8.6 Hz, 1H), 6.81 (dd, J = 8.7, 2.7Hz, 1H), 6.69 (dd, J = 13.3, 8.4 Hz, 2H), 6.49 (d, J = 2.6 Hz, 1H),5.58-5.08 (m, 2H), 3.59 (s, 3H), 2.74 (t, J = 6.8 Hz, 2H), 1.83 (s, 3H),1.78 (t, J = 6.7 Hz, 2H), 1.32 (d, J = 1.6 Hz, 6H). 506.3 [M + H]⁺ +++

¹H NMR (400 MHz, Chloroform-d) δ 8.49-8.29 (m, 2H), 8.04 (dd, J = 2.1,1.5 Hz, 1H), 7.31-7.27 (m, 2H), 7.04-6.94 (m, 2H), 6.92- 6.71 (m, 2H),2.76 (t, J = 6.8 Hz, 2H), 1.88 (s, 3H), 1.79 (t, J = 6.7 Hz, 2H), 1.32(s, 6H). 487.3 [M + H]⁺ ++

¹H NMR (400 MHz, Chloroform-d) δ 8.01-7.96 (m, 1H), 7.55-7.48 (m, 1H),7.24-7.20 (m, 2H), 7.05- 7.00 (m, 1H), 7.01-6.92 (m, 1H), 6.85-6.78 (m,1H), 6.73 (d, J = 8.5 Hz, 1H), 3.96 (d, J = 0.7 Hz, 3H), 2.75 (dd, J =7.2, 6.2 Hz, 2H), 2.41 (t, J = 0.7 Hz, 3H), 1.84 (s, 3H), 1.78 (t, J =6.7 Hz, 2H), 1.32 (d, J = 0.7 Hz, 6H). 506.3 [M + H]⁺ +++

¹H NMR (400 MHz, chloroform-d) δ 7.58-7.51 (m, 2H), 7.47-7.41 (m, 2H),7.22-7.16 (m, 2H), 7.03 (d, J = 2.4 Hz, 1H), 6.96 (dd, J = 8.6, 2.5 Hz,1H), 6.85-6.80 (m, 1H), 6.70 (d, J = 8.6 Hz, 1H), 5.16 (s, 2H), 2.73 (t,J = 6.7 Hz, 2H), 1.83 (s, 3H), 1.77 (t, J = 6.7 Hz, 2H), 1.30 (d, J =2.0 Hz, 6H) 516.5 +++

¹H NMR (400 MHz, chloroform-d) δ 7.58-7.52 (m, 2H), 7.44-7.39 (m, 2H),7.21-7.16 (m, 2H), 7.03 (d, J = 2.4 Hz, 1H), 6.98-6.93 (m, 1H),6.85-6.80 (m, 1H), 6.70 (d, J = 8.6 Hz, 1H), 5.13 (s, 2H), 3.61 (s, 3H),2.73 (t, J = 6.7 Hz, 2H), 1.82 (s, 3H), 1.76 (t, J = 6.7 Hz, 2H), 1.30(d, J = 1.8 Hz, 6H). 530.5 +++ +, 20000 nM ≥ A₂ ≥ 500 nM; ++, 500 nM >A₂ ≥ 100 nM; +++, 100 nM > A₂.

BIOLOGICAL EXAMPLES Measuring Efficacy of Chemokine Modulators

Examples of In Vitro Assays—Reagents

MOLT-4 cells were obtained from the American Type Culture Collection(Manassas, Va.) and cultured in RPMI tissue culture medium supplementedwith 10% fetal calf serum (FCS) in a humidified 5% CO₂ incubator at 37°C. Recombinant human chemokine proteins TECK was obtained from R&DSystems (Minneapolis, Minn.). ChemoTX™ chemotaxis microchambers werepurchased from Neuro Probe (Gaithersburg, Md.). CyQUANT™ cellproliferation kits were purchased from Molecular Probes (Eugene, Oreg.).Calcium indicator dye Fluo-4 AM was purchased from Molecular Devices(Mountain View, Calif.).

In Vitro Assays

A variety of assays can be used to evaluate the compounds providedherein, including signaling assays, chemotaxis (migration assays),ligand binding assays, and other assays of cellular response. Chemokinereceptor signaling assays can be used to measure the ability of acompound, such as a potential CCR(9) antagonist, to block CCR(9) ligand-(e.g. TECK)-induced signaling. Blocking such signaling can be useful intreating various diseases such as inflammatory bowel diseases, anallergic disease, psoriasis, atopic dermatitis, asthma, fibroticdiseases, graft rejection, immune mediated food allergies, autoimmunediseases, Celiac disease, rheumatoid arthritis, thymoma, thymiccarcinoma, leukemia, solid tumor, acute lymphocytic leukemia, melanoma,primary sclerosing cholangitis, hepatitis or post-operative ileus.

Chemotaxis assays can also be used to assess receptor function andevaluate the compounds provided herein. These assays are based on thefunctional migration of cells in vitro or in vivo induced by an agent,and can be used to assess the binding and/or effect on chemotaxis ofligands, inhibitors, or agonists. A variety of chemotaxis assays areknown in the art, and any suitable assay can be used to evaluate thecompounds of the present disclosure. Examples of suitable assays includethose described in PCT/US97/15915; Springer et al., WO 94/20142; Bermanet al., Immunol. Invest., 17:625-677 (1988); and Kavanaugh et al., J.Immunol., 146:4149-4156 (1991)).

Evaluation of a Test Modulator in a Serum Chemotaxis Assay

A serum chemotaxis assay was used to determine the efficacy of potentialreceptor antagonists at blocking the migration mediated throughchemokine receptors, such as CCR(9). This assay was performed using theChemoTX® microchamber system with a 5-nm pore-sized polycarbonatemembrane. MOLT-4 cells were collected by centrifugation at 400×g at roomtemperature, then suspended at 50 million/ml in human serum, containing50 mM HEPES (final pH of 7.2). The compound being tested or anequivalent volume of its solvent (DMSO) was then added to the cell/serummixture at a final DMSO concentration of 0.125% (v/v), and this mixturewas then incubated together at 37° C. for one hour. Separately,recombinant human TECK was diluted with chemotaxis buffer (HBSS+0.1%BSA), generally spanning a range from 0.1 nM to 500 nM, after which 29μl of diluted chemokine was placed in the lower wells of the ChemoTX®plate. The 5-nm (pore size) polycarbonate membrane was placed onto theplate, and 20 μL of the cell/compound mixture was transferred onto eachwell of the membrane. The plates were incubated at 37° C. for 90minutes, after which the polycarbonate membranes were removed and 5 μlof the DNA-intercalating agent CyQUANT (Invitrogen, Carlsbad, Calif.)was added to the lower wells. The amount of fluorescence, correspondingto the number of migrated cells, was measured using a Spectrafluor Plusplate reader (TECAN, San Jose, Calif.).

The A2 values were calculated from the following equation, comparing theefficacy of the test compound with that of the DMSO-only control atequi-active chemokine levels:

Log(A ₂)=log[drug(M)]−log[(A′/A)−1]

where A reflects the potency of the agonist in the absence of antagonistand A′ reflects the potency of the agonist in the presence of antagonistat a given concentration of drug ([drug(M)]).

In Vivo Efficacy Models for Human IBD

T cell infiltration into the small intestine and colon have been linkedto the pathogenesis of human inflammatory bowel diseases which includeCoeliac disease, Crohn's disease and ulcerative colitis. Blockingtrafficking of relevant T cell populations to the intestine is believedto be an effective approach to treat human IBD. CCR(9) is expressed ongut-homing T cells in peripheral blood, elevated in patients with smallbowel inflammation such as Crohn's disease and Coeliac disease. CCR(9)ligand TECK is expressed in the small intestine. It is thus believedthat this ligand-receptor pair plays a role in IBD development bymediating migration of T cells to the intestine. Several animal modelsexist and can be used for evaluating compounds of interest, such aspotential CCR(9) antagonists, for an ability to affect such T cellmigration and/or condition or disease, which might allow efficacypredictions of antagonists in humans.

Animal Models with Pathology Similar to Human Ulcerative Colitis

A murine model described by Panwala and coworkers (Panwala et al., JImmunol., 161(10):5733-44 (1998)) involves genetic deletion of themurine multi-drug resistant gene (MDR). MDR knockout mice (MDR−/−) aresusceptible to developing a severe, spontaneous intestinal inflammationwhen maintained under specific pathogen-free facility conditions. Theintestinal inflammation seen in MDR−/− mice has a pathology similar tothat of human inflammatory bowel disease (IBD) and is defined by Th1type T cells infiltration into the lamina propria of the largeintestine.

Another murine model was described by Davidson et al., J Exp Med.,184(1):241-51(1986). In this model, the murine IL-10 gene was deletedand mice rendered deficient in the production of interleukin 10(IL-10−/−). These mice develop a chronic inflammatory bowel disease(IBD) that predominates in the colon and shares histopathologicalfeatures with human IBD.

Another murine model for IBD has been described by Powrie et al., Int.Immunol., 5(11):1461-71 (1993), in which a subset of CD4+ T cells(called CD45RB(high)) from immunocompetent mice are purified andadoptively transferred into immunodeficient mice (such as C.B-17 scidmice). The animal restored with the CD45RBhighCD4+ T cell populationdeveloped a lethal wasting disease with severe mononuclear cellinfiltrates in the colon, pathologically similar with human IBD.

The TNF ARE(−/−) Model.

The role of TNF in Crohn's disease in human has been demonstrated morerecently by success of treatment using anti-TNF alpha antibody by Targanet al., N. Engl. J Med., 337(15):1029-35 (1997). Mice with aberrantproduction of TNF-alpha due to genetic alteration in the TNF gene(ARE−/−) develop Crohn's-like inflammatory bowel diseases (seeKontoyiannis et al., Immunity, 10(3):387-98 (1999)).

Examples of In Vivo Efficacy Assays

Evaluation of a Test Modulator in a CCR(9) Dependent T Cell TraffickingModel

Single cell suspensions were prepared from spleens and lymph nodes ofOT-I Tg CD45.1 mice. 15×10⁶ total cells (about 3×10⁶ CD8 T cells) wereinjected into sex-matched congenic CD45.2 C57BL/6n mice (8-10 weeksold). 24 hours later, animals were immunized via oral gavage with 25 mgOvalbumin protein (Sigma-Aldrich, St. Louis, Mo.)+10 ug Cholera Toxin(Calbiochem, San Diego, Calif.). CCR(9) antagonist compound 1.063(Table 1) was administered prior to oral ovalbumin in a time framedictated by their mouse pharmacokinetics and dosed throughout. Five dayspost immunization, animals were euthanized, and small intestines wereharvested. Peyer's patches were removed and, after flushing with PBS,the gut was opened on a wet square of Optima fabric (AllegianceHealthcare). The mucosa was scraped with a scalpel and then dissociatedby stirring in 50 ml of medium containing 10% newborn calf serum and DTT(1 mM) for 15 min at room temperature. After centrifugation, pelletswere resuspended in PBS containing 10% newborn calf serum, vortexed for3 min, and rapidly passed through a glass wool column (1.6 g packed in a20-ml syringe; Fisher Scientific). IEL were further purified on aFicoll-Paque gradient and stained with mAbs for flow cytometry analysis.Transferred OT-1 Tg CD45.1 T cells were detected and quantified by flowcytometry. In this model treatment with a compound of the disclosureresulted in a significant reduction in the frequency of OT-1 Tg CD45.1 Tcells that traffic to the small intestine in response to antigen.

Evaluation of a Test Modulator in a Model of Inhibition of HIV Spread

In the bone marrow/liver/thymus, or “BLT” mouse, non-obese diabetic(NOD)/SCID mice (which lack endogenous T and B cells) are surgicallyimplanted with fetal thymic and liver organoids, as in the SCID-husystem. The mice are then sub-lethally irradiated and transplanted withautologous CD34⁺ stem cells obtained from fetal liver which take upresidence in the murine bone marrow, effectively receiving a human bonemarrow transplant and resulting in a range of human cells in peripheralblood, including mature T and B lymphocytes, monocytes, macrophages, anddendritic cells, all of which show extensive infiltration of organs andtissues including liver, lung, and gastrointestinal tract. Followingtransplantation, a compound of the disclosure is administered totransplanted mice to inhibit the trafficking of human cells to thegastrointestinal tract, a major source of T cell/HIV interaction.Compound efficacy is measured as a reduction in blood viral load bystandard techniques.

Evaluation of a Test Modulator in a Model of Arthritis

A 17-day study of type II collagen-induced arthritis is conducted toevaluate the effects of a modulator on arthritis-induced clinical ankleswelling. Rat collagen-induced arthritis is an experimental model ofpolyarthritis that has been widely used for preclinical testing ofnumerous anti-arthritic agents (see Trentham et al., J. Exp. Med.146(3):857-868 (1977), Bendele et al., Toxicologic Pathol. 27:134-142(1999), Bendele et al., Arthritis. Rheum. 42:498-506 (1999)). Thehallmarks of this model are reliable onset and progression of robust,easily measurable polyarticular inflammation, marked cartilagedestruction in association with pannus formation and mild to moderatebone resorption and periosteal bone proliferation.

Female Lewis rats (approximately 0.2 kilograms) are anesthetized withisoflurane and injected with Freund's Incomplete Adjuvant containing 2mg/mL bovine type II collagen at the base of the tail and two sites onthe back on days 0 and 6 of this 17-day study. The test modulator isdosed daily by sub-cutaneous injection from day 9 to day 17 at a dose of100 mg/kg and a volume of 1 mL/kg in the following vehicle (24.5Cremaphore EL, 24.5% common oil, 1% Benzylalcohol and 50% Distilledwater). Caliper measurements of the ankle joint diameter are takendaily, and reducing joint swelling is taken as a measure of efficacy.

Evaluation of a Test Modulator in a Mouse Model of Asthma

This example describes a procedure to evaluate the efficacy ofantagonists for treatment of asthma. An animal model of asthma can beinduced by sensitizing rodents to an experimental antigen (e.g. OVA) bystandard immunization, and subsequently introducing that same antigeninto the rodents lung by aerosolization. Three series of rodent groups,comprising 10 rodents per group, are actively sensitized on Day 0 by asingle i.p. injection with 100 ug OVA in phosphate-buffered saline(PBS), along with an adjuvant e.g. aluminum hydroxide. At 11 days aftersensitization, the animals are placed in a Plexiglas chamber andchallenged with aerosolized OVA (1%) for 30 minutes using the ultrasonicnebulizer (De Vilbliss). One series of mice additionally receives PBSand Tween 0.5% i.p. at the initial sensitization, and at differentdosing schedules thereafter, up until the aerosolized OVA challenge. Asecond series consists of groups of mice receiving different doses ofthe CCR4 antagonist given either intraperitoneally, intra-venously,sub-cutaneously, intra-muscularly, orally, or via any other mode ofadministration at the initial sensitization, and at different dosingschedules thereafter, up until the aerosolized OVA challenge. A thirdseries of mice, serving as positive control, consists of groups treatedwith either mouse IL-10 i.p., anti-IL4 antibodies i.p., or anti-IL5antibodies i.p. at the initial sensitization, and at different dosingschedules thereafter, up until the aerosolized OV A challenge. Animalsare subsequently analyzed at different time points after the aerosolizedOVA challenge for pulmonary function, cellular infiltrates inbronchoalveolar lavage (BAL), histological examination of lungs, andmeasurement of serum OVA specific IgE titers.

It is therefore intended that the foregoing detailed description beregarded as illustrative rather than limiting, and that it be understoodthat it is the following claims, including all equivalents, that areintended to define the spirit and scope of this disclosure.

1.-37. (canceled)
 38. A compound or a pharmaceutically acceptable saltthereof, selected from Table 1 and having activity of ++ or +++. 39.-44.(canceled)
 45. A compound selected from:


46. The compound of claim 45, which is:


47. The compound of claim 45, which is:


48. The compound of claim 45, which is: